摘要
我们分离到了一株产生分泌型青霉素G酰化酶的巨大芽孢杆菌(Bacillus megateriumBM1)。用pBR322作载体,将该菌的青霉素G酰化酶基因克降到大肠杆菌(Escherichia coliMC1061)中,得到含有9.9kb插入片段的重组质粒pBmPA4。分析了该质粒的限制酶酶切图谱,并经体外缺失获得含4.9kb插入片段的质粒pBmPA5。pBmPA4和pBmPA5在E.coliMC1061中均能表达,表达受苯乙酸诱导。
A strain of Bacillus meqaterium (designated BM1) that producespenicillin G acylase secretorily has been isolated and identified. Itsstructural gene has been cloned in E. coli MC1061 using the vectorpBR322.A penicillin G acylase positive clone, pBmPA4, was screenedout by the NIPAB test paper method. The cloned DNA fragment is 9.9kb made of two (2.1kb+7.8kb) EcoRI fragments, and its restrictionmap has been established. A clone pBmPA5, containing 4.9 kb insertedDNA in the vector pWR13, has been developed by in vitro deletingpBmPA4. pBmPA5 still contains the full lenghth of penicillin G acylasegene. Both pBmPA4 and pBmPA5 strains can express penicillin Gacylase genes in the host strain E.coli MC1061. Penicillin G acylaseproduced by the clone strains is under the induction of phenylacetic acid
出处
《生物工程学报》
CAS
CSCD
北大核心
1991年第1期47-53,共7页
Chinese Journal of Biotechnology
关键词
PGA
巨大芽孢杆菌
基因克隆
表达
Penicillin G acylase
gene cloning
expression
Bacillus meqaterium
