Barletta R G, Snapper B, Cirillo J D, Connell N D, Kim D D, Jacobs W R, Bloom B R
Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461.
Res Microbiol. 1990 Sep-Oct;141(7-8):931-9. doi: 10.1016/0923-2508(90)90132-a.
Bacille Calmette-Guerin (BCG), currently the most widely used vaccine in the world, was originally administered for many years as an oral vaccine. The low frequency of serious complications, inexpensive production, and adjuvanticity make BCG an ideal candidate for a recombinant vaccine vehicle. Although mycobacteria are slow growing and not yet well characterized genetically, we have recently developed technology for the genetic manipulation of BCG and other mycobacteria. Phage and plasmid systems based on a shuttle strategy to manipulate DNA in Escherichia coli and transfer it to mycobacteria have been developed. We have established that the aminoglycoside phosphotransferase gene can be used as an effective selectable marker in the mycobacteria and that a foreign antigen from Mycobacterium leprae can be expressed in BCG. Furthermore, a thorough analysis of mycobacterial expression sequences has been undertaken to optimize the expression of foreign antigens in BCG. We constructed an expression probe shuttle plasmid with beta-galactosidase as reporter gene, and have used it successfully to identify multiple mycobacteriophage DNA sequences with varying levels of constitutive or regulable promoter activity. Further genetic advances required for development of recombinant BCG into an effective recombinant vaccine vehicle, including possibilities for oral administration, are adumbrated.
卡介苗（BCG）是目前世界上使用最广泛的疫苗，最初多年来一直作为口服疫苗使用。严重并发症发生率低、生产成本低廉以及具有佐剂性，使得卡介苗成为重组疫苗载体的理想选择。尽管分枝杆菌生长缓慢且尚未在基因方面得到充分表征，但我们最近已开发出用于卡介苗和其他分枝杆菌基因操作的技术。基于穿梭策略在大肠杆菌中操纵DNA并将其转移至分枝杆菌的噬菌体和质粒系统已被开发出来。我们已经确定氨基糖苷磷酸转移酶基因可作为分枝杆菌中的有效选择标记，并且来自麻风分枝杆菌的一种外源抗原可在卡介苗中表达。此外，已对分枝杆菌表达序列进行了全面分析，以优化外源抗原在卡介苗中的表达。我们构建了以β-半乳糖苷酶作为报告基因的表达探针穿梭质粒，并已成功用其鉴定出具有不同水平组成型或可调控启动子活性的多个分枝杆菌噬菌体DNA序列。将重组卡介苗开发成有效的重组疫苗载体所需的进一步基因进展，包括口服给药的可能性，已作了概述。
