畜牧兽医学报2017，Vol.48Issue(9)：1579-1590,12.DOI:10.11843/j.issn.0366-6964.2017.09.003
猪ADAM10基因编码区及其剪接体克隆、序列分析与组织表达研究
Coding Sequence Cloning, Alternative Splicing, Sequence Analysis and Tissue Expression of Porcine ADAM10 Gene
摘要
Abstract
The objective of this study was to clone the coding sequence (CDS) of porcine ADAM10 gene and its splicing variant,analyze the gene structure and functions using bioinformatics methods,and investigate the mRNA levels of ADAM10 gene and its splicing variant in different tissues in porcine.According to the sequence of porcine ADAM10 gene in GenBank,the specific primers were designed for cloning the CDS of ADAM10 gene and its splicing variant using reverse transcription-PCR (RT-PCR) and T/A cloning methods.The protein structure of ADAM10 and its splicing variant were analyzed through bioinformatics methods.The mRNA expression patterns of porcine ADAM10 gene and its splicing variant in different tissues in porcine were analyzed by semi-quantitative PCR.The results showed that the complete CDS of ADAM10 gene was 2 247 bp,encoding 748 amino acids.The CDS of splicing variant,missing the sequence of exon 2-7 and partial exon 8,had a length of 1 344 bp which encoded 447 amino acids.Compared with the intact protein,the splicing variant protein missed the leading peptide sequence.However,the splicing variant protein contained signal peptide,transmembrane domain and metalloproteinase and depolymerized domains.Futhermore,the tertiary structures of the 2 proteins were identical.Sequence alignment and phylogenetic analysis showed that the intact ADAM10 protein was highly conserved among species.Semi-quantitative PCR analysis showed that the intact mRNA of ADAM10 gene was highly expressed in spleen and lowly in kidney,mammary gland,leg muscle,fallopian tube,ovary,uterus and small intestine;the mRNA of splicing variant was highly expressed in lung and lowly in spleen and stomach.Our study provides the basic materials for further studying on the structure and biological function of porcine ADAM10 gene.关键词
猪/ADAM10基因/克隆/剪接体/组织表达Key words
pig/ADAM10 gene/cloning/splicing variant/tissue expression分类
农业科技引用本文复制引用
陈闻达,裴悦,周晓龙,赵阿勇,杨松柏..猪ADAM10基因编码区及其剪接体克隆、序列分析与组织表达研究[J].畜牧兽医学报,2017,48(9):1579-1590,12.基金项目
国家自然科学基金(31501921) （31501921）
浙江省自然科学基金(LQ15C170001) （LQ15C170001）
浙江省农业(畜禽)新品种选育重大科技专项(2016C02054-3) （畜禽）
