抑制性消减杂交技术在细菌活的非可培养状态研究中的应用

Application by suppression subtractive hybridization in the study on viable but nonculturable state bacteria

在基因分离与克隆中,构建各种cDNA文库是近些年在发现新基因及研究基因功能常用的基础工具之一。其中,抑制性消减杂交技术是一种高效分离差异表达基因的方法,在分离筛选细菌活的非可培养状态与正常菌株差异基因中,应用这种研究方法是研究细菌VBNC状态株毒力、致病性和耐药性及菌株遗传分化关系的重要途径。本文旨在介绍SSH构建消减cDNA文库在原核生物中的应用及对其在细菌VBNC状态新基因筛选中的应用前景进行综合论述,并对其进一步研究提出了展望。

cDNA library constructions are basic tools of detecting new gene and re- searching gene function in gene separation and cloning in recent years. Suppression Subtractive Hybridization is one of the most efficient method in isolating differen- tially expressed genes between viable but non-culturable state bacteria and normal strains, SSH is the important way in studying virulence, pathogenicity, drug resis- tance and genetic differentiation relations of strains in VBNC bacteria. The objec- tive of this paper was to discuss the construction strategies of cDNA by SSH in pro- karyotic organism and its application to isolate differentially expressed genes in VBNC bacteria, and it put forward the outlook for further research.