1) Depends on protein. But yea, uniprot is usually good. Also, you likely wont be adding it into the bacterial genome but rather a plasmid.
On Tuesday, March 22, 2016 at 10:02:21 PM UTC-7, Justin S wrote:
-- 2) There are *lots* of cloning techniques. I personally like using some vague cloning techniques, but basic ones include classic restriction enzyme cloning and gibson assembly. You wouldn't use CRISPR for this cloning.
3) Western blot
However, all these steps can variate depending on what exactly you want to express. If its just overexpression of a bacterial protein, just grab a kit to do so.
-Koeng
On Tuesday, March 22, 2016 at 10:02:21 PM UTC-7, Justin S wrote:
Hi all,I'm a total beginner, but I want to get started doing some experimental biology of my own. I'm hoping the community here can help fill in some gaps for me!1) Suppose I want to modify the DNA of some bacteria with the goal of producing some protein. How does one go about figuring out what sequence needs to be added to the bacterial genome?2) How is the sequence inserted? I'm aware of crispr, but I'm guessing there are other techniques. What's available to the layperson?3) How does one verify the desired proteins are produced?
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