Joseph Joan, Fernández-Lloris Raquel, Pezzat Elías, Saubi Narcís, Cardona Pere-Joan, Mothe Beatriz, Gatell Josep Maria
AIDS Research Unit, Hospital Clínic/IDIBAPS-HIVACAT, University of Barcelona, 08036, Barcelona, Spain.
J Biomed Biotechnol. 2010;2010:357370. doi: 10.1155/2010/357370. Epub 2010 Jun 27.
Mycobacterium bovis Bacillus Calmette-Guérin (BCG) as a live vector of recombinant bacterial vaccine is a promising system to be used. In this study, we evaluate the disrupted expression of heterologous HIV-1gp120 gene in BCG Pasteur host strain using replicative vectors pMV261 and pJH222. pJH222 carries a lysine complementing gene in BCG lysine auxotrophs. The HIV-1 gp120 gene expression was regulated by BCG hsp60 promoter (in plasmid pMV261) and Mycobacteria spp. alpha-antigen promoter (in plasmid pJH222). Among 14 rBCG:HIV-1gp120 (pMV261) colonies screened, 12 showed a partial deletion and two showed a complete deletion. However, deletion was not observed in all 10 rBCG:HIV-1gp120 (pJH222) colonies screened. In this study, we demonstrated that E. coli/Mycobacterial expression vectors bearing a weak promoter and lysine complementing gene in a recombinant lysine auxotroph of BCG could prevent genetic rearrangements and disruption of HIV 1gp120 gene expression, a key issue for engineering Mycobacterial based vaccine vectors.
牛分枝杆菌卡介苗（BCG）作为重组细菌疫苗的活载体是一种很有前景的应用系统。在本研究中，我们使用复制型载体pMV261和pJH222评估了异源HIV-1 gp120基因在卡介苗巴斯德宿主菌株中的表达缺失情况。pJH222在卡介苗赖氨酸营养缺陷型菌株中携带一个赖氨酸互补基因。HIV-1 gp120基因的表达由卡介苗hsp60启动子（在质粒pMV261中）和分枝杆菌属α-抗原启动子（在质粒pJH222中）调控。在筛选的14个rBCG:HIV-1 gp120（pMV261）菌落中，12个显示部分缺失，2个显示完全缺失。然而，在筛选的所有10个rBCG:HIV-1 gp120（pJH222）菌落中均未观察到缺失。在本研究中，我们证明了在卡介苗重组赖氨酸营养缺陷型菌株中携带弱启动子和赖氨酸互补基因的大肠杆菌/分枝杆菌表达载体可以防止基因重排和HIV 1gp120基因表达的缺失，这是构建基于分枝杆菌的疫苗载体的关键问题。
