Slauch J M, Mahan M J, Mekalanos J J
Harvard Medical School, Boston, MA.
Biotechniques. 1994 Apr;16(4):641-4.
In order to understand the genetic regulation of bacterial genes whose products are important for pathogenesis, one needs to measure the expression of the genes during the infection process. We have devised a method to measure the transcriptional activity of such genes from bacteria recovered directly from infected host tissue. Starting with bacterial strains containing lacZ transcriptional fusions to the genes of interest, animals can be infected, with subsequent isolation of infected host tissue. Here we describe the separation of bacterial cells away from a particular host tissue and the subsequent measurement of the activity of beta-galactosidase, the product of the lacZ gene, in the bacterial cells. This assay is sensitive enough to compensate for the potentially low number of bacteria recovered from the infection site.
为了了解其产物对致病作用很重要的细菌基因的遗传调控，需要在感染过程中测量这些基因的表达。我们设计了一种方法来测量直接从受感染宿主组织中回收的细菌中此类基因的转录活性。从含有与感兴趣基因的lacZ转录融合体的细菌菌株开始，可以感染动物，随后分离受感染的宿主组织。在这里，我们描述了从特定宿主组织中分离细菌细胞以及随后测量细菌细胞中lacZ基因产物β-半乳糖苷酶的活性。该测定法足够灵敏，能够弥补从感染部位回收的细菌数量可能较低的情况。
