Romero D A, Klaenhammer T R
Department of Microbiology, North Carolina State University, Raleigh 27695-7624.
Appl Environ Microbiol. 1992 Feb;58(2):699-702. doi: 10.1128/aem.58.2.699-702.1992.
The lactococcal insertion sequence IS946 was used to construct suicide vectors for insertion of heterologous DNA into chromosomal and plasmid sequences of Lactococcus lactis subsp. lactis. Electroporation of L. lactis strains, including the recombination-deficient strain MMS362, with the suicide vector pTRK145 yielded 10(1) to 10(3) transformants per micrograms of DNA. pTRK145 insertions occurred primarily in the chromosome, with one insertion detected in a resident plasmid. Vector-specific probes identified junction fragments that varied among transformants, indicating random insertions of pTRK145.
乳球菌插入序列IS946被用于构建自杀载体，以便将异源DNA插入乳酸乳球菌乳亚种的染色体和质粒序列中。用自杀载体pTRK145对包括重组缺陷菌株MMS362在内的乳酸乳球菌菌株进行电穿孔，每微克DNA可产生10(1)至10(3)个转化体。pTRK145的插入主要发生在染色体上，在一个常驻质粒中检测到一个插入。载体特异性探针鉴定出在转化体之间不同的连接片段，表明pTRK145的随机插入。
