RACE法克隆巴什拜羊SP-A基因及序列分析  

RACE Cloning and Sequential Analysis of Pulmonary Surfactant-associated Protein A in Bashibay Sheep

作　　者：王继雪 孙延鸣[1] WANG Ji-xue;SUN Yan-ming(College of Animal Science and Technology,Shihezi University,Xinjiang Shihezi 832003,China)

机构地区：[1]石河子大学动物科技学院,新疆石河子832003

出　　处：《西南农业学报》2018年第10期2196-2201,共6页Southwest China Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31460686)

摘　　要：【目的】为扩增巴什拜羊的肺表面活性物质相关蛋白A(Pulmonary surfactant-associated protein A,SP-A)基因cDNA全长序列并对该序列进行分析。【方法】根据已报道的GenBank中绵羊SP-A基因的开放阅读框序列来设计RACE引物,后用RACE法对巴什拜羊SP-A基因的3'端和5'端序列进行扩增,经华大公司测序和DNAMAN软件拼接最终得到巴什拜羊的SP-A基因cDNA序列全长。【结果】克隆得到SP-A基因的cDNA其全长为1962 bp,其开放阅读框序列长度为789 bp,编码的氨基酸数量为262,经DNASTAR软件分析其蛋白的分子量为27.53 kDa,等电点4.949。分析系统进化树后发现巴什拜羊与绵羊氨基酸的序列同源性最高。【结论】本研究结果可为后续更加深入研究新疆巴什拜羊SP-A基因的功能奠定基础。【Objective 】This paper aimed to clone the full length of pulmonary surfactant-associated protein A in Bashibay sheep and analyse its sequence.【Method】RACE primers according to the SP-A sequence of GenBank in sheep was designed,the sequence of the 3＇ and 5＇ ends of the SP-A gene by using RACE method was cloned,and the full length of SP-A gene of Bashibay sheep by sequencing and splicing was got.【Result】The full length of SP-A gene of Bashibay sheep with an open reading frame of 789 bp was 1962 bp and encode 262 amino acids.The molecular weight of the protein encoded by the gene was 27.53 kDa and its isoelectric point was 4.949.The analysis of system evolution indicated that the amino acid sequence of Bashibay sheep and sheep was the most homologous.【Conclusion】The results of this study lay a foundation for the further study of the SP-A gene function.

关 键 词：SP-A 巴什拜羊 CDNA末端快速扩增 序列分析 

分 类 号：S826[农业科学—畜牧学]