摘要
以含有P19和cyt1A基因的以色列亚种72MD质粒的97kbHindⅢ片段为模板进行PCR扩增,分别获得P19基因和cyt1A基因片段。与表达载体pUHE24连接转化大肠杆菌XL1,获得3个克隆株。LZ19含有P19基因;pLZcyt1A含有cyt1A基因;LZ19A含有P19和cyt1A两个基因。利用cyt1A蛋白质可使大肠杆菌细胞致死的特性,在IPTG诱导下,测定了各克隆基因表达对大肠杆菌有致死作用;pLZ19A对大肠杆菌的起始致死作用明显快于pLZcyt1A,这种现象可能是P19基因促进cyt1A基因高表达的结果。
The P19 gene and cyt1 A gene were obtained by PCR with 9 7kb HindIII fragment containing P19 gene and cyt 1A gene from 72MD plasmid of Bt as a template.Digested PCR producs were ligated to expression vector pUHE24 and transformed into E.coli XL 1.Three clones,LZ19 harboring P19 gene,LZcyt1A harboring cyt 1A gene,LZ19A harboring P19 gene and cyt1 A gene,were screened.The growth cure of cloned strains were determined under IPTG indution.The result showed that pLZ19 did not affect the growth of E.coli ,pLZcyt1A is typically Lethal for E.coli, Lethal initial effeciency of pLZ19A is much higher than that of pLZcyt1A.Probably,this was a result that P19 gene enhanced initial expression of cyt1 A gene in E.coli .
出处
《微生物学报》
CAS
CSCD
北大核心
1999年第2期114-119,共6页
Acta Microbiologica Sinica