Cloneliness is Next to Godliness

September 9, 2011

Rheanna Sand

And now... part 3 of 3 of the most nail-biting, heart-in-your-throat molecular biology story ever published on this website!


Hello again, folks. Ready for more science? REAL science? Okay!


After dealing with the Wrath of Kits last week, and hitting several stupid roadblocks (which were cleared when I realized a solution had been made improperly), I found some success in Playing God. I created mutants! Three to be exact. Two at position 308 and one at position 329 in the protein called mouse Kv1.2. The next step after creating the mutant DNA is creating the mutant protein. That requires the use of a live animal cell - one of the largest cells in the whole animal kingdom, the Xenopus laevis oocyte (egg). Xenopus is a totally aquatic clawed frog that lives in Africa, and has the incredibly convenient feature of being full of eggs almost all year. So, our department has a colony, and we have a tank of females that we use simply for their eggs. And yes, I have to open them up to get them. The surgery takes about 20 minutes once they are under, and after I suture them up, they take about an hour to wake up before I return them to the tank.


See? All better. The eggs must then be rinsed to get rid of all the yolk (as seen in the picture below) and digested with enzymes to get all the connective tissue out of the way.



After a few more rinses, some sorting, and treatment with another solution, they are ready for injection. That is, if the mRNA worked... which is where I left off last time! Long story short, YES the mRNA worked:



But when I injected them into the eggs, two looked like this:


But one of them looked like THIS:


And preliminary results suggest it is less affected by the snail toxin than the original channel. Which means, I might be on the right track. Which means... you just might read about this in a scientific journal someday.


Hope you have enjoyed this dose of real science! It's what I do every day and although it probably sounds like a different language, I'm always trying to find ways to talk about it so people understand why I spend so much time in the lab. BECAUSE IT'S COOL!



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