We custom-designed talen activator sequence and it is in a vector. I need to express this protein and need to isolate it. I do not know the nature of protein.
We have pet 45b expression vector t7 inducing competent cell for expression.
I need suggestions regarding the problem and caution along with expert advice and if possible some protocols for expression of talen (even a good pdf may be useful).
There is usually no need to purify TALEN proteins in order to use them. you simply clone them in the appropriate vector and transfect the vector into your target cells. although, if you need to express and purify them you can do the same as you do with any other proteins. but first, you must make sure that your custom designed TALEN does not bind to any sequence in your bacterial host's genome other wise it would induce double stranded breaks in the host's genome which may lead to loss of a large population of your expressing cells and dramatically decrease the amount of protein expressed.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology