[Research of a screening strategy for high prokaryotic expression clone].
The low expression in E. coli can be often explained by the beginning sequence that causes low translational initiation. A simple selection procedure is then used to enrich those sequences from the bank that lead to high levels of translation. The selection procedure is based on the use of a translationally coupled tetracycline resistance gene. The basic steps are as follows: (1) Construction of selection vector pBV223; (2) Screening high expression clones through different tetracycline concentrations. Several clones have been selected in 60 microg/ml concentration of tetracycline. So this system will provide an important way in preparation of some cytokines which have a low expression level in E. coli but have significant economic value.
Li
F
F
Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052.
Li
Y
Y
Shao
H
H
Zhang
Z
Z
Hou
Y
Y
D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005821:Genetic Techniques; D005822:Genetic Vectors / Q000235:genetics; D013752:Tetracycline / Q000494:pharmacology
D013752:Tetracycline