Molecular Characterization of Heterologous HIV-1gp120 Gene Expression Disruption in Mycobacterium bovis BCG Host Strain: A Critical Issue for Engineering Mycobacterial Based-Vaccine Vectors
Molecular Characterization of Heterologous HIV-1gp120 Gene Expression Disruption in Mycobacterium bovis BCG Host Strain: A Critical Issue for Engineering Mycobacterial Based-Vaccine Vectors
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DOI：
10.1155/2010/357370
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发表时间：
2010-01-01
影响因子：
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通讯作者：
Maria Gatell, Josep
中科院分区：
文献类型：
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作者：
Joseph, Joan;Fernandez-Lloris, Raquel;Maria Gatell, Josep
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应用前景的系统。本研究利用复制型载体pMV 261和pJH 222对异源HIV-1gp 120基因在BCG巴斯德宿主菌中的表达进行了研究。pJH 222携带BCG赖氨酸营养缺陷型中的赖氨酸互补基因。HIV-1 gp 120基因的表达受BCG hsp 60启动子（pMV 261）和分枝杆菌（Mycobacteria spp.）α-抗原启动子（在质粒pJH 222中）。在筛选的14个rBCG：HIV-1gp 120（pMV 261）克隆中，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-1gp 120（pJH 222）菌落中均未观察到缺失。在这项研究中，我们证明了E。在重组卡介苗赖氨酸营养缺陷型细胞中构建含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 1gp 120基因表达的破坏，这是工程化分枝杆菌疫苗载体的关键问题。
Mycobacterium bovis Bacillus Calmette-Guerin (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.