藏绵羊胃溶菌酶基因的克隆、定量分析及其蛋白性质和结构的预测  

Sequence Analysis and Detection of mRNA Expression Levels of Stomach Lysozyme Gene in Tibetan Sheep(Ovis aries)

作　　者：朱莲莲[1] 江明锋[1,2] 张鹏[1] 骆美蓉[1] 李建波[1] 任洪辉 王永[1] 

机构地区：[1]青藏高原动物遗传资源保护与利用四川省重点实验室、西南民族大学生命科学与技术学院,四川成都610041 [2]国家民委与教育部动物遗传育种重点实验室,四川成都610041

出　　处：《中国畜牧杂志》2013年第23期12-17,共6页Chinese Journal of Animal Science

基　　金：国家科技支撑计划课题(2012BAD13B06);四川省科技创新产业链示范工程项目(2011NZ0003);中央高校基本科研业务费专项资金(11ZYXS25,10NZYZJ01)

摘　　要：本实验旨在研究藏绵羊溶菌酶组织基因组成、分布情况和蛋白质结构。通过RT-PCR方法克隆测序得到藏绵羊溶菌酶基因核苷酸序列,并对氨基酸的组成和功能活性位点如53-谷氨酸和71-天冬氨酸等进行统计;运用生物信息学软件对藏绵羊溶菌酶的等电点、信号肽序列等性质进行分析,并构建系统进化树;运用荧光定量PCR方法对藏绵羊的瘤胃、瓣胃、皱胃、肾脏、气管和肝脏分别进行检测溶菌酶基因的部分组织分布情况及其mRNA的表达水平。结果表明:此绵羊溶菌酶基因核苷酸序列为447 bp,编码148个氨基酸残基;其mRNA的表达水平在藏绵羊的瘤胃和瓣胃相对较高,与其他4种组织比较差异显著(P<0.05);皱胃与肾脏也有显著性差异(P<0.05)。This experiment was designed to study the gene composition, tissue distribution and protein structure of Tibetan sheep lysozyme. The nucleotide sequence of the lysozyme gene in Tibetan sheep was sequenced and cloned by RT-PCR, and bioinformatics softwares was used for its analysis, isoelectric point, signal peptide sequence, amino acid composition and functional activty sites, such as 53-Glu , 71-Asp, etc., was predicted; also phylogenetic tree was constructed by comparing with properties of several other lysozyme genes; then different expression levels of the gene in Tibetan sheep rumen, omasum, abomasum, kidney, trachea and liver in tissue were detected by fluorescence quantitative PCR. The results indicated that the length of nucleotide sequence in Tibetan sheep lysozyme gene was 447 bp, encoding 148 amino acid residues; its mRNA relatively expressed higher levels in Tibetan sheep＇s rumen and omasum, significantly different from other four organizations （P〈0.05）; There were also significant differences between abomasum and kidneys （P〈0.05）.

关 键 词：溶菌酶 克隆 MRNA表达 藏绵羊 

分 类 号：S826.2[农业科学—畜牧学]