荣昌猪唾液腺全长cDNA文库的构建及ESTs分析

发布时间：2019-01-10 13:48

【摘要】：【目的】构建荣昌猪唾液腺全长cDNA文库,并进行部分表达序列标签(ESTs)的测序及分析,为荣昌猪种质资源的保存、开发和挖掘提供重要的分子信息。【方法】以14日龄荣昌猪仔猪为材料,取其唾液腺,采用SMART法构建唾液腺全长cDNA文库,并进行ESTs测序、注释及功能分析。【结果】初级文库滴度为5.88×106PFU/mL,重组率为99.67%,大部分插入片段长度为400~2 000bp。共获得144条有效ESTs序列,共计108条非重复序列,包括23个重叠群和85个单一序列,77(占71.29%)条序列为已知功能基因,31(占28.71%)条为未知功能序列,其中18条为新基因。GO分类和KEGG途径分析结果显示,荣昌猪唾液腺表达基因功能主要包括细胞成分、分子功能和生物学过程3个类群。在细胞成分类群中,细胞外的成分所占比例最高;在分子功能类群中,酶抑制剂活性所占比例最高;而在生物过程类群中,以转运和定位等过程最突出。表达的基因主要参与补足物和凝固级联、百日咳、加压素调控的水再吸收及味觉传导等代谢途径。【结论】成功构建了荣昌猪唾液腺全长cDNA文库,解析了荣昌猪唾液腺的表达特征。
[Abstract]:[objective] to construct the full-length cDNA library of Rongchang pig salivary gland, and to sequence and analyze the partial expression sequence tag (ESTs) for the preservation of Rongchang pig germplasm resources. To develop and excavate the important molecular information. [methods] the salivary glands of 14 day old Rongchang piglets were taken as materials, the full-length cDNA library of salivary glands was constructed by SMART method, and ESTs sequencing was performed. Results the titer of the primary library was 5.88 脳 10 ~ (6) PFU / mL, the recombination rate was 99.67, and the length of most of the inserted fragments was 400 ~ 2 000 BP. A total of 144-active ESTs sequences were obtained, including 23 overlapping groups and 85 single sequences. 77 (71.29%) were known functional genes, 31 (28.71%) were unknown functional sequences. The results of GO classification and KEGG pathway analysis showed that the gene function of salivary gland in Rongchang pig was mainly composed of three groups: cell component, molecular function and biological process. The proportion of extracellular components was the highest in the cell classification group, the proportion of enzyme inhibitor activity was the highest in the molecular functional group, while in the biological process group, transport and localization were the most prominent processes. The expressed genes are mainly involved in the metabolic pathways of complement and coagulation cascade, pertussis, vasopressin regulated water reabsorption and taste conduction. [conclusion] the full-length cDNA library of Rongchang pig salivary gland was successfully constructed. The expression characteristics of salivary glands in Rongchang pig were analyzed.
【作者单位】：西南大学荣昌校区动物医学系;
【基金】：国家自然科学基金项目(31201807)
【分类号】：S828

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