多浪羊IL-1β基因的克隆及序列分析

Cloning and Sequence Analysis of IL- 1β from Duolang Sheep in Xinjiang

【目的】进行多浪羊IL-1β基因的克隆及序列分析,为多浪羊IL-1β的功能研究及多浪羊免疫应答基因的筛选奠定基础。【方法】根据GenBank中NM_001009465.2的mRNA序列设计引物,以新疆多浪羊淋巴细胞的总RNA为模板,用RT-PCR扩增多浪羊IL-1β基因序列。【结果】IL-1β基因序列包含了一个862 bp的开放阅读框,成熟蛋白编码的氨基酸序列长度为286个氨基酸。通过IL-1-β核苷酸多序列比对发现,第70、163、180、269位上四个相对保守的碱基处出现了四个突变;通过IL-1-β氨基酸多序列比对,第24、36位上两个氨基酸发生突变。通过GenBank中的Blastn序列比较分析,多浪羊的IL-1β基因与普通绵羊的同源性高达99%。【结论】在进化树体系中,多浪羊与反刍动物集成一束,它们与其它哺乳动物源于共同的祖先,但分别处于进化树上不同的分支。

[Objective]The purpose of this project aims to clone and analyse the IL-1β gene in order to lay a foundation for genetic screening of Duolang sheep.[Method] The study was based on the primers from genebank and the total RNA was extracted from Duolang sheep lymphocytes,and then the IL-1 βby was amplified by RT-PCR.After sequencing the IL-1β,the gene contained an ORF of 801 bp,and the length of amino acid mature protein coding.[Result] Four relative conserved bases were found at the position of 70,163,180,269 which mutations occurred by multiple sequence alignment; the two amino acid mutations occurred at the position of 24 and 36 after multiple amino acid alignment; as a result,we know that the homology between Duolang sheep and other ordinary sheep is up to 99%.[Conclusion] The evolution reviews that Duolang sheep and the other mammals are originated from the same ancestor,but located at the different branches of the evolutionary tree.