Satchidanandam Vijaya, Amara Rama Rao, Uchil Pradeep Devappa, Singh Varsha
Department of Microbiology and Cell Biology, Indian Institute of Science, Room 254A, Sir C.V. Raman Avenue, 560012, Bangalore, India.
FEMS Microbiol Lett. 2003 Jan 28;218(2):365-70. doi: 10.1016/S0378-1097(02)01185-0.
We report efficient expression of the Mycobacterium tuberculosis gene Rv3881c in Escherichia coli from its M. tuberculosis promoter, attributable to an E. coli consensus Pribnow box and ribosome binding site. The N-terminal sequence of the recombinant E. coli-generated protein was identical to the predicted open reading frame of Rv3881c and transcription of the Rv3881c gene initiated at the same nucleotide position in both bacteria. We demonstrate the utility of this promoter for rapid analysis of expression in E. coli of heterologous gene constructs, for subsequent expression from the genomes of slow-growing mycobacteria such as Mycobacterium bovis-BCG. M. tuberculosis Rv3881c homologues were present in other pathogenic mycobacteria such as M. bovis-BCG, Mycobacterium szulgai and Mycobacterium kansasii.
我们报道了结核分枝杆菌基因Rv3881c在大肠杆菌中从其结核分枝杆菌启动子高效表达,这归因于大肠杆菌共有普里布诺框和核糖体结合位点。重组大肠杆菌产生的蛋白质的N端序列与Rv3881c预测的开放阅读框相同,并且Rv3881c基因在两种细菌中的转录起始于相同的核苷酸位置。我们证明了该启动子可用于快速分析异源基因构建体在大肠杆菌中的表达,以便随后从生长缓慢的分枝杆菌如牛分枝杆菌卡介苗的基因组中表达。结核分枝杆菌Rv3881c同源物存在于其他致病性分枝杆菌中,如牛分枝杆菌卡介苗、苏尔加分枝杆菌和堪萨斯分枝杆菌。