Optimising expression of the large dynamic range FRET pair mNeonGreen and superfolder mTurquoise2^ox for use in the Escherichia coli cytoplasm.

ABSTRACT: The fluorescent proteins superfolder mTurquoise2^ox (sfTq2^ox) and mNeonGreen function excellently in mammalian cells, but are not well expressed in E. coli when forming the N-terminus of constructs. Expression was increased by decreasing structures at the start of their coding sequences in the mRNA. Unfortunately, the expression of mNeonGreen started from methionine at position ten as optimisation introduced an alternative RBS in the GFP N-terminus of mNeonGreen. The original start-codon was not deleted, which caused the expression of isomers starting at the original start-codon and at the start-codon at the beginning of the GFP N-terminus. By omitting the GFP N-terminus of mNeonGreen and optimising the structure of its mRNA, the expression of a mixture of isomers was

SUBMITTER: Mertens LMY 

PROVIDER: S-EPMC9606377 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature