Expression of recombinant genes in Esherichia coli - promoted by high levels of arginine-incorporating transfer RNA

摘要

Recombinant genes contg. AGA and/or AGG codons for Arg are expressed in E. coli by transformation of the E. coli cells with an expression vector contg. the recombinant gene, provided that the amt. of Arg-incorporating tRNA recognising the AGA and/or AGG codons (hereafter called Arg tRNA) present in the cells is increased by a factor of at least 5 over the amt. normally present. Also claimed are (a) expression vectors contg. a recombinant gene and a gene coding for Arg tRNA, and (b) expression-assisting vectors which contain and express a gene coding for Arg tRNA and are compatible with an expression vector having a ColE1 replication origin. USE/ADVANTAGE - The process is esp. useful for promoting expression of eukaryotic genes having a large no. of AGA and/or AGG codons, e.g. those coding for human urokinase, t-PA, HIV proteins (e.g. gp.41 or p24) or yeast alpha-glucosidase. High levels of Arg tRNA in the transformed cells increases the proportion of recombinant protein produced and improves the vitality of the cells.