长白猪CIDEC基因、启动子的克隆及其在不同组织中的表达研究  

Cloning and Expression of CIDEC Gene and Its Promotor from Landrace

作　　者：李红强 郭振清 孙健 LI Hongqiang;GUO Zhenqing;SUN Jian(College of Agriculture and Biotechnology,Hebei Normal University of Science&Technology,Hebei Qinhuangdao,066600,China)

机构地区：[1]河北科技师范学院农学与生物科技学院,河北秦皇岛066600

出　　处：《中国畜牧杂志》2020年第6期55-60,168,共7页Chinese Journal of Animal Science

基　　金：国家自然科学基金资助项目(31702101);河北省高等学校科学研究青年基金项目(QN2016136);河北省人才培养工程资助项目(A201901055)。

摘　　要：本研究旨在克隆长白猪诱导细胞凋亡DFF45样效应因子C(CIDEC)基因及其转录起始位点前约2000bp启动子区域,并检测其在长白猪不同组织中的表达水平,从而为研究CIDEC在猪中的作用及表达调控机制提供依据。选择8头6月龄去势长白猪,提取肝脏RNA,利用RT-PCR技术扩增CIDEC基因的CDS序列,使用生物信息学相关软件分析其结构和功能;屠宰后采集长白猪心脏、肝脏、脾脏、肺、肾脏、胃、皮下脂肪、腿肌、颈阔肌、肱二头肌、小肠、空肠等组织样品,分别提取各样品的总RNA,利用qRT-PCR检测CIDEC在各组织中的mRNA表达量;提取长白猪肝脏总DNA,采用降落PCR方法获得CIDEC启动子序列,利用Methprimer在线软件对CpG岛区域进行预测。结果表明:克隆得到的长白猪CIDEC基因CDS序列长度为747bp,编码238个氨基酸,进化关系分析发现猪与羊、牛的亲缘性较近。CIDEC基因在长白猪的各个组织均有表达,其中在皮下脂肪组织表达量最高,在肾脏中表达量最低。对CpG岛区域预测结果显示CIDEC基因CDS序列不存在典型的CpG岛。The aim is to clone the CIDEC gene and its promoter region about 2 000 bp before transcription start site from landrace, and to detect its expression level in different tissues of Landrace, so as to provide a basis for studying the role of CIDEC and its expression regulation mechanism. In this paper, 8 Landraces aged 6 months were taken as objects. Liver RNA was extracted and the CDS sequences of CIDEC were obtained by RT-PCR, and the structure and function of CIDEC were analyzed by bioinformatics software. Tissue samples of heart, liver, spleen, lung, kidney, stomach, subcutaneous fat, leg muscle, platysma, biceps brachii, small intestine and jejunum were collected after slaughter. The total RNA was extracted from each tissue sample, and the mRNA expression of CIDEC was detected by qRT-PCR(quantitative real-time PCR). The total DNA from the liver of landrace was extracted and the CIDEC promoter sequence was obtained by descending PCR. The results showed that the CDS sequence length of the cloned CIDEC gene was 747 bp and encoded 238 amino acids, and the evolutionary relationship showed that pigs were closely related to sheep and cattle. The CIDEC gene is expressed in all tissues of landrace, with the highest expression in subcutaneous adipose tissue and the lowest expression in kidney. Subsequently, the promoter sequence of CIDEC gene of landrace was cloned, and the CpG island region was predicted using the online software Methprimer. The results showed that there was no typical CpG island in this sequence.

关 键 词：CIDEC 基因克隆 降落PCR 长白猪 

分 类 号：S828.2[农业科学—畜牧学]