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          Characterization of Brassica rapa RAP2.4-Related Proteins in Stress Response and as CUL3-Dependent E3 Ligase Substrates.      Comment   Translate Page      
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Characterization of Brassica rapa RAP2.4-Related Proteins in Stress Response and as CUL3-Dependent E3 Ligase Substrates.

Cells. 2019 Apr 10;8(4):

Authors: Mooney S, Al-Saharin R, Choi CM, Tucker K, Beathard C, Hellmann HA

The turnip Brassica rapa has important economic value and represents a good model system to study gene function in crop plants. ERF/AP2 transcription factors are a major group of proteins that are often involved in regulating stress-responses and developmental programs. Some ERF/AP2 proteins are targets of CULLIN3-based E3 ligases that use BTB/POZ-MATH proteins as substrate receptors. These receptors bind the transcription factor and facilitate their ubiquitylation and subsequent degradation via the 26S proteasome. Here, we show tissue and stress-dependent expression patterns for three Brassica rapa ERF/AP2 proteins that are closely related to Arabidopsis thaliana AtRAP2.4. Cloning of the Brassica genes showed that the corresponding proteins can assemble with a BPM protein and CULLIN3, and that they are instable in a 26S proteasome dependent manner. This work demonstrates the conserved nature of the ERF/AP2-CULLIN3-based E3 ligase interplay, and represents a first step to analyze their function in a commercially relevant crop plant.

PMID: 30974760 [PubMed]

          7 Best Free Linux Disk Cloning Software      Comment   Translate Page      

One of the key responsibilities of a system administrator is to ensure the safety of an organisation’s computer data. This is a critical task for any organisation, given that the loss of important data could have profound implications on its future prosperity.

There are a number of different strategies that a system administrator will need to use to ensure that data is preserved. For example, the person will need to maintain the computer system and/or network, by applying operating system updates and patches, by performing regular backups, and ensuring that the system is kept secure at all times. Disk cloning is a highly popular method of performing comprehensive backups of the operating system, installed software, and an organisation’s data with the minimum of effort.

However, disk cloning software is not simply helpful for system backups. It has a wide range of other uses such as provisioning new computers in the workplace, restoring computers from a master image, and system recovery.

To provide an insight into the quality of software that is available, we have compiled a list of 7 high quality Linux disk cloning software. Hopefully, there will be something of interest for anyone who needs to perform system admin tasks.

Read more

          Open Source for Advanced Beginners #1: bashplotlib      Comment   Translate Page      

This is a project for advanced beginner programmers.

The goal of this project is to help you make the leap from small practice exercises to complex, thorny programs like what you might encounter in your first future programming job. I’ve found an accessible open source library and have written 7 detailed feature requests for it. Your task is to read the library’s code, understand it, and write these features. You’ll get plenty of guidance along the way, but not too much, and you’ll have to do all the difficult bits yourself. I’ve written more about why and how I think this project will help you go from Advanced Beginner to Beginning Advanced here.

We’re going to be working with bashplotlib, a Python library that displays ASCII graphs in the terminal. Created by Greg Lamp, it is small, approachable, and sensibly-written, whilst still having plenty of room for improvement in both its code and functionality.

We’re going to make bashplotlib’s graphs look more attractive; add automated unit tests; and even add an entirely new graph type. We’ll learn how to search through a new codebase; when and when not to worry about code duplication; and how to safely change the way that a program is structured. After each task I’ve written pull requests (PRs) on GitHub to show you what I would do. You can use these PRs to help you out if you get stuck and to compare notes once you’re done.

Before we begin, let’s do some quick setup.


I’ve “forked” (git’s word for “made my own copy of”) the bashplotlib repo and put it on my GitHub account. I did this so that no one else can sneak in and implement the 7 features in this project before you’ve had a chance to. By the time you read this, the code in the main bashplotlib repo may have changed significantly, but my fork will stay exactly the same forever.

Here’s how to get bashplotlib set up on your computer.

Task A: Fork your own version of bashplotlib on GitHub

(optional but recommended)

I recommend that you fork your own version of my fork. This will allow you to practice using git to push your work back up to GitHub, and will also allow you to show the world what you’ve been working on. If you can’t or don’t want to do this then you can skip to step 3 and clone my version of bashplotlib directly. Don’t worry about breaking anything; GitHub wouldn’t let you mess up my version even if you wanted to. To fork your own bashplotlib repo:

  1. If you don’t have a GitHub account, create one. Follow GitHub’s instructions for linking your computer to your account. When following the instructions, note that we’re going to do our cloning using HTTPS, not SSH.
  2. Go to and click “Fork”. This will create your own copy of the bashplotlib repo.

Task B: Clone bashplotlib

To clone bashplotlib and copy the files onto your computer:

  1. In your terminal, cd to the directory you want to work in (eg. cd ~/Documents/Programming)
  2. If you forked your own version of bashplotlib in the previous section, clone it by running the command:
    git clone$YOUR_GITHUB_NAME/bashplotlib.git
    If you didn't fork your own version, clone my version by running:
    git clone
    If you can't get cloning to work, despite having wasted your whole goddamn afternoon trying, you can download the code as a standard zip file by clicking on the Clone or download button on

Task C: Set up a scratch file

The rigorous best practice for working with Python libraries is to use a tool called virtualenv. If you want to try setting it up then you can follow the instructions here, but for now I recommend saving yourself the potential frustration and heartache of installing a new tool and following these simpler and perfectly valid steps instead:

  1. In your terminal, cd into the new directory you just created by your clone operation (eg. cd ./bashplotlib)
  2. Make a scratch Python file. The name isn't important; is fine.
  3. Copy this code into it:
    from bashplotlib.scatterplot import plot_scatter
    x_coords = [-10,20,30]
    y_coords = [-10,20,30]
    width = 10
    char = 'x'
    color = 'default'
    title = 'My Test Graph'
  4. Run this code with the command python ./ If the command prints a scatterplot then your setup is complete and you're ready to take on the world. If something goes wrong then congratulations, you get to practice your debugging skills to try and figure out what. Send me a message if you get stuck.

Let’s start small and get warmed up.

1. Make the title box more stylish

bashplotlib displays its graph titles in a moderately nice-looking box:


Let’s make these titles a bit more eye-catching. The way that the box edges smash into each other in the corners is too harsh. Let’s replace the corners with plus-signs so that the lines merge nicely with each other instead.


Open bashplotlib/ and have a look around. Where is the code that prints the title box?

Once you’ve found the box-printing code, you’ll see that it uses a function that is defined in another file in the bashplotlib project. Which file? Use your text editor’s “Find in project” (or similar) feature, and locate the function’s code. (Hint - if you want to find where a Python function is defined, search for "def my_function_name" instead of just my_function_name)

Once you believe you’ve found the box-generating function, make sure that you really have found it by changing some of the function’s code. For example, change the function so that the sides of the box are displayed using the ^ character instead of the |. Run your scratch file (cd to the root of the bashplotlib project and run python ./, and make sure that the output is what you expect to be. If it is, you know you’re in the right place. If it isn’t, work out why.

Now that you’ve found the piece of code that builds the title box, you’re ready to change it so that it puts plus-signs in the box’s corners. Go ahead.

Make sure that you don’t accidentally break anything in the process. For example, make sure that the title box stays the same width as the graph, and that the text inside the box stays centered. If we had unit tests then we wouldn’t have to worry about accidentally breaking anything, because our tests would tell us if we had. We’re going to write some unit tests in section 4 of this project, but unfortunately we don’t have any yet. For now you’ll just have to be extra careful and manually check your work.

How I did it

Here’s my pull request.


Done it? Congratulations! Keep going; add further embellishments as your personal taste dictates. For example:

  • Work out how to left- and right-align the title text. Allow users to choose between left-, right-, and centre-align by passing in a title_align parameter to the plot_scatter function.
  • If the title is longer than the width of the graph, truncate it and add ellipses () instead of overflowing.
  • Add 2 layers of edging, like so:

  • Use your imagination! Make the title as elegant and beautiful as you can.

2. Label the axes

bashplotlib’s graphs are very clear, but the axes don’t have titles. This makes it hard to know what a graph is trying to tell you. Does that y-axis represent GDP growth or the amount of carbon it the atmosphere? Are the units percent, parts-per-million, or years-until-extinction?


Let’s add the ability to optionally pass in titles for the x- and y-axes of scatterplots, and display these titles like so:

In addition, let’s add --xtitle and --ytitle options to the command line interface at the bottom of the file so that the command line tool can use them too.


Feel free to tidy up the code a little if it helps you understand it. I personally felt like len(get_scale(xs, False, size) was being written out too many times, so I assigned its result to a variable at the top of the _plot_scatter function and used that throughout the rest of the function.

How I did it

Here’s my pull request.


Save space and show off your skills by printing the y-axis title vertically down the side of the graph, like so:

3. Show the 0-axes in the middle of the graph

The README for bashplotlib shows an example scatterplot that has its “0-axes” (lines showing where the axes cross 0) displayed in the middle of the graph:

However, it seems that somewhere the code that creates these 0-axes got lost (nb. I did not delete the code for teaching purposes, it really does seem to have got lost).


0-axes can be very useful for interpreting graphs, so let’s add them back in. Here’s how our 0-axes should work:

  • If our data points cross the x-axis, we should print a 0-axis for the x-axis (eg. (-10, 10), (-5, 15), (0, 20), (5, 25)). If they cross the y-axis, we should print a 0-axis for the y-axis too (eg. (-10, -10), (-5, -5), (0, 0), (5, 5))
  • We should only print a 0-axis if our data points cross 0 for an axis. If our data doesn’t cross 0, we shouldn’t print a 0-axis
  • If we print 0-axes for both the x- and y-axes, we should print an O where they cross
  • If there is a data point on top of an axis line, we should make sure to print the data point and not the axis line


When I was working on this feature myself, I realized that I had to actually work out what the double-nested-for-loop inside the _plot_scatter function was doing. Up until this point I hadn’t needed to, so I hadn’t bothered. I added a few comments and tweaked the code slightly to make it easier for me to understand. Feel free to do this yourself if it helps you.

How I did it

Here’s my pull request.


Allow users to hide these new axes if they pass an optional hide_zero_axes argument into the plot_scatter function. 0-axes should be displayed by default, so the default value for this argument should be False.

4. Make the library “testable” and write some tests

Making changes to any codebase, especially a large, unfamiliar one, can be unnerving. You may have successfully added your new feature, but how do you know that it works correctly with every possible edge-case? And how do you know that you haven’t inadvertently broken an already existing feature in some subtle (or not-so-subtle) but important way?

The slow, unscalable, error-prone way to test your code is to manually run it using lots of different combinations of parameters, and laboriously check that everything looks correct. This is better than not testing at all, but does nothing to prevent someone else from unwittingly breaking your new feature in the future.

The solution is “unit tests”. A unit test is a block of code that calls functions from your main code, and verifies that these functions behave correctly. For example, if you wanted to check the correctness of a new function called sum_digits, which takes a number and returns the sum of its digits (in base 10), one of your tests might be:

result = sum_digits(543)
expected_result = 12

if result == expected_result:

You might also write other tests that make sure that sum_digits works correctly for other types of number, like negative numbers, decimals, and the number 0.

Unit tests make verifying the correctness of your code much quicker and more reliable - all you have to do is run your test file and make sure none of the tests fail. It also means that future people who work with your code (including yourself) will know how it’s meant to behave, and will be less likely to accidentally break it.

At Stripe, where I work, I would guess that the codebase has well over 10,000 tests, and maybe many more. This means that anyone can add new features and update existing ones, without being unduly concerned that they will unwittingly destroy the company.

Making bashplotlib “testable”

bashplotlib does not currently have any tests. This is a perfectly reasonable choice by the author. Writing and maintaining tests takes time, and he probably felt that the library was small and experimental enough that he didn’t want the bother. Nonetheless, adding tests would make the library more reliable and easier to work with. While I was writing this project I noticed and fixed a small bug that would have been caught automatically if the library had had any tests.

However, right now bashplotlib is not very “testable”. A piece of code’s “testability” is how easy it is to write tests for. There are many reasons why a piece of code might or might not be testable. bashplotlib’s main problem is that it prints all of its output to the terminal piece-by-piece. Automated tests for the current version of bashplotlib would have to somehow read in output that had previously been printed to the terminal. There are ways to do this (or something close to it), but they are somewhat complicated and fiddly. There is a better solution.


In this task we’ll first make bashplotlib testable, and then we’ll write tests for it.

We’ll make bashplotlib “testable” by changing the _plot_scatter function so that it no longer prints the graph, but instead builds and returns the graph as a long string. The plot_scatter function (which calls _plot_scatter) will then be responsible for printing this long string to the terminal:

def _plot_scatter(...arguments, etc...)
    graph = ""
    # a graph...
    graph += "+---+etc"
    # the rest of the graph...
    return graph

def plot_scatter(...arguments, etc...)
    # ...process arguments...
    graph_text = _plot_scatter(...arguments, etc...)

This seemingly minor shuffling of responsibilities makes the _plot_scatter function much easier to test. Now we can write test code that calls _plot_scatter, stores the result in a variable, and compares this result to a pre-defined, expected value. For example, in a new file called, we can write:

from bashplotlib import _plot_scatter

# Copy and paste the value for `expected_result`
# from terminal output that you know is correct.
expected_result = """+--------...
# Call `_plot_scatter` with the arguments that
# you know will produce the `expected_result`
# above.
result = _plot_scatter(...arguments, etc...)

if expected_result == result:

Think of as many edge-cases as you can, and write individual tests for them all.


I suggest that you tackle this task using the following steps:

  • Change _plot_scatter so that it returns a string representing the graph, and doesn’t print anything
  • Change plot_scatter so that it stores and prints the output of _plot_scatter
  • Make a new file called in the root of the library (alongside
  • Write a test for _plot_scatter to make sure that it generates a normal graph correctly. Run it using python ./
  • Think of as many edge-cases as you can, and write tests for them. Do your 0-axes display properly? What if a point is on top of a 0-axis? What happens if your title is longer than the width of the graph? Is this what you want to happen? What if you have 2 data points that are the same? Almost the same?

Your tests might have trouble interpreting color printing. Feel free to get rid of the color-printing functionality for now, or look at my version for suggestions on how to deal with it.

For another example of unit testing, see section 3 of Programming Projects for Advanced Beginners #2: Game of Life.

How I did it

Here’s my pull request.


Research PyUnit, a Python testing framework, and rewrite our tests with it. Although our hand-crafted if-statements are perfectly serviceable, most test suites are written using a testing framework (like PyUnit), which makes writing, running, and debugging tests much easier.

5. Make histograms more stylish too

So far all of our work has been on improving scatterplots. But bashplotlib can plot histograms too. Technically they’re not actually histograms; the y-axis in bashplotlib’s graphs represents a raw count, whereas in a histogram the y-axis represents “probability density”. “Bar charts” would be a more accurate word for what bashplotlib produces. But I think everyone still understands what the library is getting at, so let’s not say anything nasty on Twitter. For consistency I’m going to keep referring to them as “histograms”.


bashplotlib’s histograms generate their titles using the same box_title function that we worked on in step 1, so they automatically benefit from our previous work. However, histograms also display a statistical summary of their data, in another box below the main graph. This box is generated by hand. We’ve already got a good way of printing text inside beautiful ASCII boxes, so let’s use this method here too. This will make our statistics look more elegant and consistent, and reduce code duplication

To work on histograms, you’ll have to call plot_hist from your file. Try to set this up yourself.


We have a small problem - box_text currently assumes that you only want to print a single line inside your box, but our statistics are spread out over multiple lines. To solve this snafu, update box_text so that the argument it accepts is a list of strings, not just a single string. Have the function display each element of the list on a separate line, and surround all the lines with an ASCII box. We’ll then use the function to create our stats box like so:

center = ... # Defined earlier in the file
summary_lines = [
    "## Summary ##".center(center),
    ("observations: %d" % n).center(center),
    # ...etc...
print(box_text(summary_lines, center))

To maintain compatibility with our existing code, update the places where box_text is used for displaying titles so that the titles are passed in as a single element list, like this:


How I did it

Here’s my pull request.


Because we’re now using the box_text function to render the histogram statistics box, we no longer get an emphasizing line in between “Summary” and “observations” (see picture above). I thought that line looked quite fetching, so let’s bring it back. Update box_text so that it takes an argument of a “list of lists”. box_text should print out each line in the sub-lists, as normal, but in between each top level list it should print an ASCII line. For example, the code:

  ["Sub-title1", Sub-title2"],
  ["Point1", "Point2", "Point3"]

should produce the output:

|   Title  |
|  Point1  |
|  Point2  |
|  Point3  |

Make sure all the other usages of box_text still work.

6. Add labels to histogram axes


Do the same thing that you did in task 2 - add optional titles to the x- and y-axes - but this time for histograms. It’s a lot easier the second time round, isn’t it?


You’ll probably find that you repeat code between the and files. Repeating code like this should always make you feel a little bit uncomfortable, and should prompt you to consider ways to reduce or eliminate the duplication. That said, sometimes repeating code can be an entirely pragmatic choice.

In our situation, we could either try to wrap up our axis title code in some sort of print_axis_title function that lives in utils/, or we could copy and paste and duplicate our code in both and For this task, I could personally go either way. On the one hand, it’s always good to reduce repeated code. But on the other hand, it’s not obvious to me that scatterplots and histograms will always and forever want to use the same logic for printing their axis labels. For example, what happens if one of them wants to print some extra characters on the same line as the axis label? We’d probably have to add something convoluted like optional extra_chars_left and extra_chars_right parameters. Trying to combine two pieces of code that might not be as combine-able as they first appear can lead to some awkward approaches. Since the code for printing axis labels is relatively simple anyway, on this occasion I lean in favor of copying and pasting.

I’m still glad that I thought about it though.

How I did it

I reckon you can tackle this one on your own.


I’ve got nothing. Call your parents and see how they’re doing.

7. Make a new graph type: horizontal histogram

This is my final feature request, and it’s a big one.


Let’s allow users to draw horizontal histograms:

This will help users display big, detailed histograms that are too tall for their short and stocky terminals, but that fit nicely when rotated.


This will be very hard. Take inspiration from, but not too much inspiration. Feel free to start from scratch if you prefer. I’d suggest that you:

  • Copy and paste into a new file
  • Make sure you can import and run its code in Test it by using it to draw some normal, still-vertical histograms
  • Think about which parts of the vertical histogram code you can reuse, and which parts will have to change. In general you’ll be able to reuse the parts of the code that deal with transforming and bucketing the data (everything above line 140 or so), since the underlying data doesn’t change. However, you will have to rewrite the parts that take the transformed data and display it as a graph (everything below line 140). For now, use copy and paste liberally and don’t worry about duplicating code.
  • Once you’ve got your horizontal histogram working, write some tests for it!

How I did it

Here’s my pull request.


Pull out as much duplicated code as you can from vertical and horizontal histograms, and move it into shared functions. These functions might perform common operations like bucketing data points and calculating scales.

8. Even more extensions

  • Calculate the “correlation” of scatter graphs using the “product moment correlation coefficient” algorithm and print it in a box below the plot. Research what these words mean. You can either look for an existing implementation of the algorithm and use that, or try to code it yourself from scratch (this involves a lot of very tricky but very interesting maths).
  • Calculate the “interquartile range” of histogram data and display it in the same box as the mean, median, standard deviation, etc. If you’re not sure what that means, look it up!
  • Use ASCII techniques to connect the dots in scatter plots. You’ll have to work out which points need to be joined together, calculate the gradient between them, and work out how best to connect them using ASCII characters.
  • Draw ASCII pie charts. Add a new bashplotlib/ file to put the logic in, and add a new pie command-line tool

That’s it

You’re done! Keep the momentum going - choose another open source project (use Google and this list for inspiration) and add experimental new features to it. Don’t worry much about actually submitting your work to the main project (unless you want to) - your goal should be to understand and experiment with new projects, not to produce something polished and perfect and ready to be deployed to production servers. Alternatively, try tackling the Programming Projects for Advanced Beginners series, where you build your own programs from scratch using a similar level of detailed (but not too detailed) guidance to that of this project.

I’d love to hear your feedback and find out how you got on. Send me an email or a Tweet (with a link to your GitHub repo if you have one) and I’ll send you back some insightful comments and a ASCII art trophy.

          Cloning 1, 2, 3      Comment   Translate Page      
Video: Cloning 1, 2, 3
Watch This Video!
Studio: Celebrity Video Distribution
Cloning 1,2,3 is a distracting, beguiling way to face our future. The hilarious prospects and zany visions help eliminate the fears and doubts. I believe the future will continue to survive will brilliant and unique discoveries to make the world stronger and more intelligent. There is no turning back. If we do not do it, someone more intelligent will. Let's have some fun and check out this whimsical method.

          Thread: Nemesis:: General:: Nemesis in Glorious Stereo Vision!!!      Comment   Translate Page      

by ravenmir

For those not familiar with Battle Systems Terrain, it's a great company in England that makes high quality cardoard terrain for gaming. They are currently fulfilling a kickstarter game called Core Space, that is a full miniatures game user their terrain.

While waiting patiently for that game I thought I'd try to build the Nemesis ship using my existing terrain sets. It was a bit tricky getting the doors and and numbering correct, but hopefully pictures will help me with future builds of this. A few of the rooms were a bit tight, so I may expand the ship slightly the next time I build it out. Battle Systems also does terrain elements, so things like their cloning chambers are perfect for the hibernation room. Can’t wait till I get the Nemesis 3D elements like eggs and dead bodies in the second wave. That will really make things pop on the board!!! They have a number of accessories that nicely compliment most the room types, computers for the comms, auto doctor tables for the surgery room, etc. It makes for a really immersive game!

          Sales Executive Trainee - Pragati Biomedical - Delhi, Delhi      Comment   Translate Page      
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          Cloning 1, 2, 3      Comment   Translate Page      
Video: Cloning 1, 2, 3
Watch This Video!
Studio: Celebrity Video Distribution
Cloning 1,2,3 is a distracting, beguiling way to face our future. The hilarious prospects and zany visions help eliminate the fears and doubts. I believe the future will continue to survive will brilliant and unique discoveries to make the world stronger and more intelligent. There is no turning back. If we do not do it, someone more intelligent will. Let's have some fun and check out this whimsical method.

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          Cloning & Modify or Request a Pokemon      Comment   Translate Page _Trainer Name_: Pain _Friend Code_: 0147 - 1463 - 9464 _Pokemon Versions_: https://www.neo...
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          (USA-MI-Grand Rapids) Research Associate-Fixed Term      Comment   Translate Page      
h3Position Summary/h3 The Williams Lab in the Department of Pediatrics and Human Development at the MSU Grand Rapids Research Center is seeking to hire a (Postdoctoral) Research Associate. The goal of our research is to determine the role of neurodevelopmental disorder candidate genes (such as those implicated in autism, intellectual disability, and epilepsy) in synapse formation and function. We specialize in the development and application of viral and transgenic tools to understand the molecular mechanisms of synapse development. Therefore, this position can offer training in diverse techniques such as cloning, biochemistry, and microscopy; or the opportunity to develop further specialized expertise in electrophysiology. h3Required Degree/h3 Doctorate -Neurology, biology, biochemistry, or a related field. h3Minimum Requirements/h3 Candidate must possess a terminal degree (PhD or equivalent) to be eligible for consideration. h3Desired Qualifications/h3 Researchers with three (3) or more years'' experience with path-clamp electrophysiology using mouse brain slice preparations are preferred. h3Required Application Materials/h3 liCV/li liCover letter/li liNames and contact information for three (3) references (references will not be contacted without first notifying the applicant)/li /ul h3Review of Applications Begins On/h3 03/26/2019 h3Summary of Health Risks/h3 Work with animals or unfixed animal tissue. h3Website/h3 http:/ h3MSU Statement/h3 Michigan State University has been advancing the common good with uncommon will for more than 160 years. One of the top research universities in the world, MSU pushes the boundaries of discovery and forges enduring partnerships to solve the most pressing global challenges while providing life-changing opportunities to a diverse and inclusive academic community through more than 200 programs of study in 17 degree-granting colleges.
          Following the Evolutionary Track of a Highly Specific L-Arginine Oxidase by Reconstruction and Biochemical Analysis of Ancestral and Native Enzymes.      Comment   Translate Page      
Related Articles

Following the Evolutionary Track of a Highly Specific L-Arginine Oxidase by Reconstruction and Biochemical Analysis of Ancestral and Native Enzymes.

Appl Environ Microbiol. 2019 Apr 12;:

Authors: Nakano S, Niwa M, Asano Y, Ito S

Following the evolutionary track of enzymes can help elucidate how enzymes attain their characteristic functions, such as thermal adaptation and substrate selectivity, during the evolutionary process. Ancestral sequence reconstruction (ASR) is effective for following evolutionary processes if sufficient sequence data are available. Selecting sequences from the data to generate a curated sequence library are necessary for the successful design of artificial proteins by ASR. In this study, we tried to follow the evolutionary track of L-arginine oxidase (AROD), an FAD-dependent amino acid oxidase (LAAO) that exhibits high specificity for L-arginine. The library was generated by selecting sequences in which the 15th, 50th, 332nd, and 580th residues are Gly, Ser, Trp, and Thr, respectively. We excluded sequences that are either extremely short or long and those with a low degree of sequence identity. Three ancestral ARODs (AncARODn0, AncARODn1, and AncARODn2) were designed using the library. Subsequently, we expressed the ancestral ARODs as well as native AROD (OkAROD) in bacteria. AncARODn0 is phylogenetically most remote from OkAROD whereas AncARODn2 is most similar to OkAROD. Thermal stability was gradually increased by extending AROD sequences back to the progenitor, while the T1/2 value of AncARODn0 was >20 °C higher than that of OkAROD. Remarkably, only AncARODn0 exhibited broad substrate selectivity similar to that of conventional promiscuous LAAO. Taken together, our findings led us to infer that AROD may have evolved from a highly thermostable and promiscuous LAAO.IMPORTANCEIn this study, we attempted to infer the molecular evolution of a recently isolated FAD-dependent L-arginine oxidase (AROD) that oxidizes L-arginine to 2-ketoarginine. Utilizing 10 candidate AROD sequences, we obtained a total of three ancestral AROD. In addition, one native AROD was obtained by cloning one of the candidate ARODs. The candidate sequences were selected utilizing a curation method defined in this study. All the ARODs were successfully expressed in E. coli for analysis of their biochemical functions. The catalytic activity of our bacterially-expressed ancestral ARODs suggests that our ASR was successful. The ancestral AROD that is phylogenetically most remote from a native AROD has the highest thermal stability and substrate promiscuity. Our findings led us to infer that AROD evolved from a highly thermostable and promiscuous LAAO. As an application, we can design artificial ARODs with improved functions compared with native ones.

PMID: 30979835 [PubMed - as supplied by publisher]

          "The Clone Wars" by TM - 4.14.19      Comment   Translate Page      
Entry Submitted by TM at 3:30 PM EDT on April 14, 2019

The better informed you are
The easier it will be for you to be able to comprehend Truths.
You will be required to come to your own conclusions because of all the falsehoods that we are constantly bombarded with.
The fact that I believe Rosenstein looks a lot like Mr Himmler. You be the judge.
I couldn't find the right pix for the Fidel Trudeau combo( Google must have removed it) but Podesta and Bennington are definitely good examples.

This journey has been long and has taken a most circuitous route. Rather convoluted some may admit to.
From crossing an ocean full of lies , deserted wastelands of emptiness and despair, down rabbit holes not fit for humanity to crawl through.
We continue to push forward.
Only the strong survive.
When in the past ,in my naïvate I made many incorrect deductions that I in my arrogance must take back.

Such as
God wouldn't allow time travel to exist...
The timelines would be compromised...
Hah ! How Naive
The gods were sleeping

There's no such things as human cloning
We, surface dwellers on this planet ,are light years away from being considered ignorant hillbillies.
I use the words surface dwellers because this world is not what it appears to be on the outside. There's an inside and also subterranean strata.
We'll leave that for a later date.
(just cause you own a smart phone doesn't make you so... Only branded)

The planet comprises of a molten center
Where all the heaviest metals pool exerting tremendous pressures... This is how gravity is created.
So we have been led to believe according to the general consensus.
The jury's out on that one
Round earth, flat earth, torus / doughnut shaped ?
I won't believe it till I see it for myself

The pyramids were built by people who had bronze tools and took those thousands of tons of sculptured granite blocks and rolled them over tree trunks...
Sooo many lies
Sooo many learned men agreed to the consensus .
I could go on and on.
Whatever for
What to believe

Within the scope of our lack of True Knowledge and Understanding we find ourselves arguing about everything under the sun as if we were experts on topics that were simply stories created for our entertainment/ enslavement.
Are We Entertained Yet?

There are people who still pay to listen to Bill & Hillary speak.
Who still worship Royals
Who Try kissing the pope's ring
My Goodness
Reality Bites
I used to believe that I possessed a grand variety of knowledge and yet; everything is suspect until it's Proven to be so.
Still we argue
Perhaps this will teach me Grace and Compassion...
The jury's still out on that one as well.

I do believe that cloning humans has been going on for a very long time now simply because we , the surface dwellers on this planet have been kept in a state of quarantine so that our development had been arrested and learning has been dripping intravenously as slowly as the GCR and planetary liberation by the Alliance

A Wild Bunch Of Full On Tards !!
This will be my last installment on
"The Clone Wars"
( spoiler alert)
We Won
Keep Your Vibrations/Frequencies High
The less you try to hang on to
The easier it is to release
All that no longer serves you
When We Ascend [Rise] We Travel Light
Release The Lies You Hang On To
Like A Baby's Blanket
There are sooo many layers of Truth to be cognizant of, that just to enumerate them as well as repeat them would take too much time; and I prefer moving forward not backwards.
This is why one must process information and let most just pass through your mind without giving it too much of your energies , simply for the sake of your own sanity.
Be careful in what you allow yourself to believe in.
Be careful in not allowing yourself to follow someone's narrative.
Be aware of talking points utilized by those who wish to decieve you.

I will utilize this young person as an example
Note to Brian.
* When a leftist NPC utilizes words and talking points that continue to repeat the same story.[ anti-Trumpian especially]
NPC is a referance to humans who are programmed. In other words they can't think for themselves.
* Trump is such a liar( can't state what he was lying about) and they become real excited about that.
* Trump puts children in cages.
* Trump is amoral and unfit for the office of Prez. ( straight from Nancy Pelosis mouth to yours)
*Trump is racist. Xenophobic
( yes dear... All leftist talking points that have been repeated ad nauseam for the last 3 years.)
* they are passionate about "Facts" that are nothing more than unproven allegations.
Wow !!
Tell me Bri ,
if one removes all these talking points... What do you have to say from Brian ?
NPC / non player character
You're just a program !!
Silly wabbitt... Twix are for kids

Note to F from Toronto
* sadly I had a young lady from Canada who went ballistic about my clone post about Justine Trudeau and pretty much proved how wrong I was because she provided so many facts about Margaret
Trudeau that even if she was in the maternity ward at that time of birth to prove Justin's birth was legitimate.
Sorry. No proof.
She simply wasn't there when the seed was planted... Sooo...
The question I had to ask was...
Who's your daddy?
And why so emotional?
You got money on this dog race that you gotta be so emotional ?

Now me
I wanted transparency but the young lady demanded privacy...
As I expressed to the reply of said email.
I will speak my mind. I will not be censored by someone who disagrees with my views.

I only post an infinitesimal amount of what I write on this site and I do so to keep myself entertained while we wait and most important to keep my frequencies in a positive vibration.
I allowed myself to become angry for quite a while by allowing the negative nellies to bring me down.
Fake pumps
Fake Intel
Ain't gonna happen no mo'
Ain't allowing Khazarian / Cabal mind manipulation.
Blame game
Everybody's fault
Waaaa !

So here is another leftist trick.
The leftist/ Bolshevik/ Khazarian/ Globalists...
I use these names because the actors are always changing names, definitions simply to confuse you.
All part of the same game.
Like a wolf in sheep's clothing.
Like calling themselves Progressive.
What they really mean is Radical
Radical Islamic
Radical Democrat
These creatures do not want anyone who disagrees with them to voice an opinion.

Anyone who comprehends the English language knows that Nationalistic feelings are patriotic feelings towards your nation.
Silly Globalists are teaching lefty kids that this means we... Conservative..
Are Nazi's...Wow.
They only see an opposing enemy. Not conservative not Republican but Right Wing as if all we want to do is go to war.
We're so bad because we want a controlled immigration policy into our sovereign nation.
Shit man. I'm Cuban born Naturalized American citizen who was brought here in 1960 at the age of two and became Naturalized when I became 18.
There's nothing wrong with following a process.
Did Obama process in 8 years any DACA. Recipients? No.
We are just bad
Bad bad citizens
So the little one's can't properly define National Socialism as what is refered to being Nazi.
Nationalism is not National Socialism
How sad is that.
( tell you a secret that's why AOC is abbreviated...Can't spell the whole name)

Wow !!
They use words like "opposed"
As if it's not a dialogue but an attack.
If my views "differ" it doesn't mean that it's in opposition of you.
Don't take it so personal
Is that how you see it?
It's really a creepy place for me when I come across these verbal manipulations
That I am becoming aware of because they leave a "slimy essence, smell, taste".
The Shite don't smell right.
In my book
People to avoid
Stories to avoid
If you get annoyed
You are an adult
Get over it

So. Let's all hold hands. Sit around the campfire. Toast some Smores and sing Cumbaya my Lord...
I forgot
You hate white men over 40 who aren't gay.[ holding hands is out]
No it's not considered rape.
Ok. No campfire. Yes
Too much carbon footprint.Wow
Can't do Smores . Right. You're vegan
And the cacao beans were not from an Organic self sustaining community approved by your handlers.
Cumbaya is out cause you hate Christians and that is a while religion that was appropriated by white people from Brown Semitic people.

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          Zanki Zero: Last Beginning - A Survival RPG from the Makers of Danganronpa | Push Square      Comment   Translate Page      
Zanki Zero is a really entertaining blend of visual novel and survival RPG gameplay. The story is just as eccentric and zany as the Danganronpa series and features that grittiness fans have come to expect. The combat system is a little simplistic, but the human cloning makes it a really unique experience. However, the title doesn't quite hit the same deliciously dark heights as Danganronpa.

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