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由于Ⅰ型人类免疫缺陷症病毒(HIV-1)通过胃肠道外及性进行传播,因此,开发HIV疫苗的主要目标是构建作用于全身和粘膜免疫系统的免疫原。为此,作者构建了一种表达重组HIV-1gp120(rgp120)的伤寒杆菌减毒株。 作者将编码rgp120的基因表达盒整合进伤寒杆菌减毒株的aroC基因座,以构建一种疫苗株。作者首先构建了双启动子系统(P_(lpp)和P_(lacUV5)),通过定位诱变将P_(1pp)的-35和-10区靠近σ70启动子共有序列,以提高蛋白表达。其次,将lacO插入启动子使lacI能调控表达。最后,在P_(1ac UVS)后加人工终止密码子,使5′到P_(lpp)核糖体结合位点(RBS)一段序列成为框内RBS,这种框内RBS-终止-RBS
Since HIV-1 is transmitted parenterally and sexually, the primary goal of developing HIV vaccines is to construct immunogens that act on the systemic and mucosal immune systems. To this end, the authors constructed a strain of Salmonella typhimurium expressing the recombinant HIV-1 gp120 (rgp120). The authors integrated the gene expression cassette encoding rgp120 into the aroC locus of the attenuated Salmonella typhimurium strain to construct a vaccine strain. The authors first constructed the dual promoter systems (P_ (lpp) and P_ (lacUV5)). The P_ (1pp) -35 and -10 regions were located close to the σ70 promoter consensus sequence by site-directed mutagenesis to enhance the protein expression. Second, inserting lacO into the promoter allows lacI to regulate its expression. Finally, an artificial stop codon is added after P_ (1 ac UVS) to make a sequence of the 5 ’to P_ (lpp) ribosome binding site (RBS) an in-frame RBS. This in-frame RBS-