细菌诱导光滑鳖甲幼虫抑制差减cDNA文库的构建与分析  

Construction and Analysis of Suppression Subtractive cDNA Library of Anatolica Polita Borealis Larva Induced by Bacteria

作　　者：刘忠渊[1] 王鹏[1] 毛新芳[1] 

机构地区：[1]新疆大学生命科学与技术学院,新疆生物资源基因工程重点实验室,新疆乌鲁木齐830046

出　　处：《新疆大学学报（自然科学版）》2016年第2期141-146,253,共6页Journal of Xinjiang University(Natural Science Edition)

基　　金：国家自然科学基金项目(31460578)资助

摘　　要：本研究以光滑鳖甲幼虫的cDNA为材料,应用抑制差减杂交（Suppression Subtractive Hybrriization,SSH）技术构建细菌诱导的光滑鳖甲幼虫抑制差减cDNA文库,并利用反向Northern杂交技术获得差异基因.结果表明,随机挑取的阳性克隆的大小均为20（0～750 bp,符合差减文库PCR产物片段的大小.通过斑点杂交技术共筛选出940个差异基因片段,对其克隆、测序及同源性分析后鉴定出70种编码蛋白的基因ESTs序列,所得ESTs序列主要涉及免疫防御、代谢过程、生长发育类等相关基因.细菌诱导光滑鳖甲幼虫抑制差减cDNA文库的构建,为进一步克隆光滑鳖甲免疫抗菌相关基因及其免疫防御机制的研究奠定了基础.In the present study,suppression subtractive cDNA library was constructed for Anatolica polita borealis larva induced by bacteria using suppression subtractive hybridization（SSH） technique,and differential genes were obtained by reverse Northern blot.The results showed that the randomly selected positive clones were all in 200-750 bp,in accord with the prediction for PCR,940 differential genes fragments were screened by dot blot hybridization.The expressed sequence tags（ESTs） sequences of 70 proteins were identified by sequence and homology analysis.The ESTs sequences were mainly involved in immune defense,metabolic processes,growing development,and other related genes.Construction of suppression subtractive cDNA library of A.polita larva induced by bacteria,for further cloning A.polita immune antibacterial related gene and its immune defense mechanism research laid the foundation.

关 键 词：光滑鳖甲 抑制差减杂交 CDNA文库 反向Northern 

分 类 号：Q523[生物学—生物化学]