摘要
用酚抽提的方法提取乳酸乳球菌的基因组DNA,利用PCR方法从乳酸菌的基因组DNA中扩增出含有苹果酸-乳酸酶基因(malolactic enzyme gene,mle)的约1.6kb的DNA片断,用1%的琼脂糖凝胶分离扩增的片断,用试剂盒回收目的基因。将回收的目的基因与pGEM-T载体连接构建mle-T载体并转化大肠杆菌DH5a,挑取阳性克隆(白色菌落),酶切鉴定并测序。SalI酶切mle-T,回收mle DNA片断,与表达载体pET-28a载体连接,构建细菌Escherichia coli表达载体。
The genomic DNA from Lactococcus lactis, lactis was extracted with the method of phenol.1.6kb malolactic enzyme gene （mle） was amplicated by PCR from the enomic DNA PCR production was purified by means of electrophoresis of 1% agrose gel and DNA reagent kit,then was linked to pGEM-T vector by T4-DNA ligase,and then transformed into E.coli DH5a,finally screening the recombinant depending on Blue-White clony.
出处
《生物技术通报》
CAS
CSCD
2006年第5期94-96,共3页
Biotechnology Bulletin
