Rapid screening for improved solubility of small human proteins produced as fusion proteins in Escherichia coli.

ABSTRACT: A prerequisite for structural genomics and related projects is to standardize the process of gene overexpression and protein solubility screening to enable automation for higher throughput. We have tested a methodology to rapidly subclone a large number of human genes and screen these for expression and protein solubility in Escherichia coli. The methodology, which can be partly automated, was used to compare the effect of six different N-terminal fusion proteins and an N-terminal 6*His tag. As a realistic test set we selected 32 potentially interesting human proteins with unknown structures and sizes suitable for NMR studies. The genes were transferred from cDNA to expression vectors using subcloning by recombination. The subcloning yield was 100% for 27 (of 32) genes for which a PCR frag

SUBMITTER: Hammarstrom M 

PROVIDER: S-EPMC2373440 | biostudies-literature | 2002 Feb

REPOSITORIES: biostudies-literature