Orientation of a DNA insert

Genetics as it applies to evolution, molecular biology, and medical aspects.

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Orientation of a DNA insert

Post by ReactUK » Sat Mar 12, 2011 12:35 pm


I've come to the fourms looking for help regarding an experiment I did recently, where I determined the orientation of a DNA insert. The experiment was a sucess and it was successfully determined.

The problem I have is trying to consoidate my knowledge on the topic I cant find any sources that actaully tell me a rife life application for this? Does knowing the oreintation of a DNA insert help in a medical field? Or with some genetics dieases? Or is it somthing completley different?!

So the question i'm asking is;

What real life applications does knowing the orientation of a DNA insert actaully help?

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Post by Darby » Sat Mar 12, 2011 1:55 pm

It seems like it would help in those areas that use them - genetic engineering of bacteria, gene therapy, production of knockout mice, etc.

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Post by JackBean » Sat Mar 12, 2011 5:26 pm

I took care of your other post...

well, usually, when you clone some gene or something, you clone only the coding sequence and the promoter is artificial already in your vector. Thus you must have it in correct orientation so that you have it upstream of your gene

Cis or trans? That's what matters.

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Post by DRT23 » Wed Mar 16, 2011 9:15 am

When you're producing a vector it's usually under your control where the cloning gene is inserted in plasmid/artificial chromosome etc. But, when you introduce a vector into the nucleus of a cell, it's usually out of control where the vector with its transgene payload is inserted in genome. (Of course, there are other applications which use uninserted, free vectors in nuclues). Thus, vector may locate in a loci that affects a native gene thus resulting in a mutation.

So, knowing the orientation of a DNA insert helps you in the applications that use genetic engineering such as GMOs, gene therapy, biotechnology etc. as Darby explains above.

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