Expression Vectors - construction (Sep/06/2005 )
i would like to know if there are any websites or any books that could give me the details for how to go about constructing a expression vector.
Look at some commercial systems (eg invitrogen, promega, ect.) that have prexisting gene expression vectors. In most cases, it can be straight forward to insert your gene of interest into a mutiple cloning site.
It depends on what organism you are using.
If you are using E. coli, you have a billion options, but I'd go with pET/gateway cloning.
If you are using B. subtilis, you are quite limited in your options. You're going to have to use an integration vector with a xylose-inducible promoter like pAX01. I've talked to several people in Kit Pagliano's lab at UCSD about the spac system and it supposedly sucks; not only is it leaky but it doesn't function well, either.
For other organisms, check the catalogs. Invitrogen makes very good stuff.
thanx for the reply, i am working on plex vector, for ecoli, where can i get info on how to go about finding the promoter and the repressor present on the vector. anyway i can find the control mechanism from the sequence i got from the invitrogen site?
Please find the details from the following link.
You can check the location of promoter, restriction sites, antibiotics resistance gene, etc here.