Suk K, Das S, Sun W, Jwang B, Barthold S W, Flavell R A, Fikrig E
Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520-8031, USA.
Proc Natl Acad Sci U S A. 1995 May 9;92(10):4269-73. doi: 10.1073/pnas.92.10.4269.
An immunological screening strategy was used to select microbial genes expressed only in the host. Differential screening of a Borrelia burgdorferi (the Lyme disease agent) expression library identified a gene (p21) encoding a 20.7-kDa antigen that reacted with antibodies in serum from actively infected mice but not serum from mice immunized with heat-killed B. burgdorferi. Selective expression of p21 in the infected host was confirmed by Northern blot analysis and RNA PCR. Further differential screening of the expression library identified at least five additional B. burgdorferi genes are selectively expressed in vivo. This screening method can be used to identify genes induced in vivo in a wide variety of pathogenic microorganisms for which a gene transfer system is not currently available.
采用免疫筛选策略来选择仅在宿主体内表达的微生物基因。对伯氏疏螺旋体(莱姆病病原体)表达文库进行差异筛选,鉴定出一个编码20.7 kDa抗原的基因(p21),该抗原能与来自活跃感染小鼠的血清中的抗体发生反应,但不能与用热灭活的伯氏疏螺旋体免疫的小鼠血清发生反应。通过Northern印迹分析和RNA PCR证实了p21在受感染宿主体内的选择性表达。对表达文库的进一步差异筛选确定了至少另外五个伯氏疏螺旋体基因在体内选择性表达。这种筛选方法可用于鉴定在多种目前尚无基因转移系统的致病微生物体内诱导表达的基因。