Cloning and expression of temperature sensitive mutant of P22 tailspike gene in Escherichia coli (Cloning of P22 tailspike gene in E. coli) = 대장균내에서 P22 tailspike 유전자의 클론닝 및 발현

Abstract

A plasmid-expression system was established in Escherichia coli for the tailspike gene (gene 9) of Salmonella phage P22 in which temperature sensitive folding mutations had been thoroughly characterized previously. Expression of the recombinant tailspike gene was under control of tac promoter and the yield of expression was 2% of total cellular protein when the the synthesis of recombinant protein was fully induced. Three mutations (ZsH304, /sUll，and 比U24)of gene 9 behaved off the E. coli plasmid exactly the same way as they did in P22-infected Salmonella; at 25 °C the mutant polypeptides fold and mature properly into native structure, while at 39 °C they could not fold but aggregate. The construction will allow a systematic approach to examine effect of amino acid substitution on protein folding and stability.