Hi colleagues! I have an Agrobacterium bacteria that contains a plasmid with the modified GFP called "mGFP5". So I want to cloning it in order to insert that gene in an expression vector in e. coli.
Nevertheless, the mgfp5 gene contains codon usage changes that disrupt a cryptic plant intron, then I wonder if that gene can be normally express in e. coli? even so containing a codon usage modified. Has someone ever worked with the mgfp5 gene to express it in e. coli?
Many thanks in advance!
It may work, but certainly you compromise the translation level because the codon usage. Unless you have very strong e coli promoter, you may not happy with the strength of signal later.
mgfp5 works fine in E. coli, I've used derivatives of this vector: https://www.addgene.org/27169/ and every time I put them into E. coli they fluoresce very well. Just keep in mind there's probably about a million E. coli-GFP expression vectors out there so depending on what you're trying to do it might be easier and cheaper just to get a premade plasmid instead of going through the cloning and verification steps.
Thanks a lot Alexandra Johnson for comment your experience and the advices as well. I will surely keep in mind about it!
Well you may be right. I thought the same before asking on this site. But anyway many thanks for your answer Delin Liang
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