匍枝根霉抑制性消减杂交差异文库的构建及分析  

Construction and Analysis of the Differences Rhizopus stolonifer cDNA Genes by Using Suppression Subtractive Hybridization

作　　者：宋哲玮 汤斌[1] 李松[1] 李祥林[1] 

机构地区：[1]安徽工程大学,微生物发酵安徽省工程技术研究中心,安徽芜湖241000

出　　处：《生物技术》2014年第2期1-5,共5页Biotechnology

基　　金：国家自然科学基金项目("匍枝根霉纤维素酶系组成解析及关键酶结构与功能研究";编号:31270315)资助~~

摘　　要：目的:研究匍枝根霉纤维素酶表达相关的调控基因。方法:使用抑制性消减杂交技术(SSH),成功构建了匍枝根霉纤维素酶系的表达差异cDNA文库。随机挑选200个阳性克隆子并进行测序,最终获得30个差异表达片段(ESTs)。结果:与GenBank比对后,获得了匍枝根霉纤维素酶表达过程中涉及的相关重要调控基因信息,包括纤维素酶系中的纤维二糖水解酶以及半纤维素酶系中的甘露糖苷酶等多种关键酶的信息。结论:为进一步研究与匍枝根霉纤维素酶表达相关的基因调控机制奠定了基础。Objective： The aim was to identify regulatory gene of cellulase expression in Rhizopus stolonifer. Method： Suppression subtrac- tive hybridization （SSH） was used in this article. A forward subtractive cDNA library was constructed successfully and positive clones from library were randomly selected to validate with enzyme digestion. Consequently, 30 expressed sequence tags （ESTs） had been found. Result：Comparing with GenBank, It indicated that cellulase expression of important regulatory genes related to Rhizopus stolonifer had been discoveried. It included cellobiohydrolase in the cellulase system, mannosidase in hemicellulase and other key enzyme informa- tion. Conclusion：The results provided the basis for future revealing the mechanism on cellulase expression.

关 键 词：纤维素酶 抑制性消减杂交技术(SSH) 匍枝根霉(Rhizopus stolonifer) 表达序列标(ESTs) 

分 类 号：Q933[生物学—微生物学]