Bacterial Transformation

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gogoplata
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Bacterial Transformation

Post by gogoplata » Sat Dec 27, 2008 5:27 am

Does anyone know how to extract DNA from white blood cells and make recombinant DNA so that the transformed bacteria will produce CD4 receptors on its cell membrane?

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MrMistery
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Post by MrMistery » Sat Dec 27, 2008 9:11 pm

you would need cDNA, and a targeting sequence for the cell membrane in bacteria. NOT as easy as it sounds...
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gogoplata
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Re: Bacterial Transformation

Post by gogoplata » Sun Dec 28, 2008 3:22 am

Thanks for the reply. I read online about cDNA, but wasn't able to find much. Could you elaborate if possible? Also, couldn't the cells be made competent by using heat shock like in normal transformation? Thanks for any help.

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Post by MrMistery » Sun Dec 28, 2008 11:33 am

well yes, but the problems you have are:
1. eukaryotic genes have introns, and bacteria can't splice out the introns. this can generally be overcome by using cDNA
2. you need to make sure your protein ends up in the plasma membrane.

But firstly, I suggest you read the details of how molecular cloning works. If you don't know what cDNA is, you're in a bit over your head.
"As a biologist, I firmly believe that when you're dead, you're dead. Except for what you live behind in history. That's the only afterlife" - J. Craig Venter

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