雌性东方蜜蜂基因表达差减文库的构建及初步分析  

Construction and Primary Analysis of Suppression Subtractive Hybridization Libraries for Female Apis cerana

作　　者：樊莹[1,2] 邹丰才[3] 董霞[1] 匡海鸥[1] 

机构地区：[1]云南农业大学东方蜜蜂研究所,云南昆明650201 [2]贵州省畜禽遗传资源管理站,贵州贵阳553000 [3]云南农业大学动物科学技术学院,云南昆明650201

出　　处：《云南农业大学学报（自然科学版）》2015年第5期710-721,共12页Journal of Yunnan Agricultural University:Natural Science

基　　金：国家自然科学基金项目(30960269)

摘　　要：蜜蜂的蜂王和工蜂是两种不同的表型,这是基于基因的差异表达,而不是遗传物质的多态性。本研究以雌性东方蜜蜂为材料,利用抑制消减杂交技术构建东方蜜蜂蜂王和工蜂的基因差异表达的文库。挑选插入片段大于200 bp的330个阳性克隆进行测序,蜂王消减文库176个,工蜂消减文库154个。测序后蜂王消减文库获得161条ESTs序列,工蜂消减文库获得142条ESTs序列。获得的序列采用DNAstar seq Man软件进行载体、接头和低质量序列去除后进行ESTs分析,蜂王消减文库获得110个contings,工蜂消减文库获得81个contings,蜂王和工蜂文库均有部分重叠的ESTs序列。在NCBI上进行在线BLASTx和t BLASTn同源比较,发现在蜂王的ESTs中有62个已知ESTs,3个未知ESTs,工蜂的ESTs中有49个已知ESTs,8个未知ESTs。通过基因本体论进行功能分析,大部分差异表达的基因为酶类和结合蛋白,主要参与基因的转录、翻译和新陈代谢过程。雌性东方蜜蜂差减文库的构建,为进一步研究蜜蜂级型分化的机制奠定了基础。The two different phenotypes of honeybees,queen and workers,are determined by the differences of gene expressions rather than the polymorphisms of the genetic material. The aim was to construct differential gene expression libraries by suppression subtractive hybridization（ SSH） for queen and worker bees of Apis cerana. We obtained 161 and 142 high quality ESTs sequences form queen and worker bees,respectively. After using DNAstar seq Man for carriers and joints removing,low quality sequence filtering,and ESTs clustering,we finally obtained 110 and 81 contings for queen and worker bees subtracted libraries, respectively. There are overlapping ESTs sequences for both queen and worker bees subtracted libraries. Using sequence homology analysis by BLASTx and BLASTn,62 out of 65 ESTs from the queen subtracted library and 49 out of 57 ESTs from the worker subtracted library are found to have homologous genes,respectively. Gene function analysis by Gene Ontology（ GO） suggested that the products of majority of the differentially expressed genes are enzymes and binding protein,which are involved in gene transcription,translation and metabolism process.The female A. cerana subtracted libraries constructed in our study provide important information for the further studies of the mechanism of caste determination.

关 键 词：东方蜜蜂 抑制消减杂交 差异表达基因 消减文库 ESTS 

分 类 号：S891.2[农业科学—特种经济动物饲养]