Luong Thanh T, Lee Chia Y
Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, 4301 W. Markham St. Mail Slot 511, Little Rock, AR 72205, United States.
J Microbiol Methods. 2007 Jul;70(1):186-90. doi: 10.1016/j.mimet.2007.04.007. Epub 2007 Apr 24.
We have previously reported the construction of Staphylococcus aureus single-copy integration vectors based on the lysogenic bacteriophage L54a site-specific recombination system. These vectors can replicate autonomously in Escherichia coli, which allows DNA manipulations. In S. aureus, the vectors, which do not possess staphylococcal replication function, can only be maintained by integrating into the chromosome. However, the original vectors have limited cloning sites and do not have protection from potential transcription of external promoters. Here we report the improved version of these vectors that circumvent these shortcomings. In addition, a second integration site based on the bacteriophage phi11 site-specific recombination system has been added such that the vectors can integrate either at the L54a attachment site or at the phi11 attachment site.
我们之前报道过基于溶原性噬菌体L54a位点特异性重组系统构建金黄色葡萄球菌单拷贝整合载体。这些载体可在大肠杆菌中自主复制，便于进行DNA操作。在金黄色葡萄球菌中，这些不具备葡萄球菌复制功能的载体只能通过整合到染色体中得以维持。然而，原始载体的克隆位点有限，且无法抵御外部启动子的潜在转录。在此，我们报道了改进后的载体版本，克服了这些缺点。此外，还添加了基于噬菌体phi11位点特异性重组系统的第二个整合位点，使得载体能够整合到L54a附着位点或phi11附着位点。
