Saviola Beatrice, Bishai William R
Basic Medical Sciences, College of Osteopathic Medicine, Western University, 309 E. Second Street, Pomona, CA 91766-1854, USA.
Nucleic Acids Res. 2004 Jan 12;32(1):e11. doi: 10.1093/nar/gnh005.
In order to create a system in which two independent plasmids can be integrated into a mycobacterial chromosome, a mycobacterial plasmid was constructed containing the phage attachment site attP from the mycobacteriophage L5 genome and additionally containing the bacterial attachment site, attB. This plasmid will integrate into the mycobacterial chromosome via recombination of the plasmid-borne attP site with the chromosomal attB site in the presence of a mycobacterial vector carrying the L5 integrase (int) gene. The integrated plasmid has a plasmid-borne attB site that is preserved and will accept the integration of additional mycobacterial plasmids containing the L5 attP site. This system should be useful in the construction of novel mycobacterial strains. In particular, this system provides a method by which several recombinant antigens or reporter constructs can be sequentially inserted into a mycobacterial strain and subsequently tested.
为了构建一个能将两个独立质粒整合到分枝杆菌染色体中的系统，构建了一种分枝杆菌质粒，该质粒含有来自分枝杆菌噬菌体L5基因组的噬菌体附着位点attP，此外还含有细菌附着位点attB。在携带L5整合酶（int）基因的分枝杆菌载体存在的情况下，该质粒将通过质粒携带的attP位点与染色体attB位点的重组整合到分枝杆菌染色体中。整合后的质粒具有一个保留的质粒携带的attB位点，并且将接受含有L5 attP位点的其他分枝杆菌质粒的整合。该系统在构建新型分枝杆菌菌株方面应该是有用的。特别是，该系统提供了一种方法，通过该方法可以将几种重组抗原或报告构建体依次插入分枝杆菌菌株中并随后进行测试。
