青海高原半细毛羊白细胞介素-8基因克隆及序列分析

Cloning and Sequence Analysis of IL-8 Gene in Qinghai Semifine-wool Sheep

本试验旨在研究青海半细毛羊白细胞介素-8(interleukin-8,IL-8)基因分子进化情况,对IL-8基因进行了克隆及序列分析。根据GenBank公布的羊IL-8基因序列设计1对特异性引物,RT-PCR扩增目的片段,将目的基因与pMD18-T克隆载体连接,转化至大肠杆菌DH5α感受态细胞,对PCR与酶切鉴定为阳性的重组质粒进行序列测定,采用生物信息学软件对IL-8基因序列进行序列分析。结果表明,羊外周血淋巴细胞中扩增出与预期大小(370bp)相符的条带,同源性分析结果显示,克隆的核苷酸序列与已知IL-8基因序列同源性为96.1%～99.0%,是较为保守的基因。

This experiment was conducted to study molecular evolution,cloning and sequence analysis of Qinghai Semifine-wool sheep interleukin-8（IL-8） gene.A pair of specific primers was designed based on GenBank.The IL-8 gene was amplified by RT-PCR and cloned into pMD18-T vector.The positive recombinant plasmids which was transformed to E.coli DH5α was identified by PCR and restriction enzyme digestion and sequenced analysis.The results showed that sheep peripheral blood lymphocytes was amplified bands consistent with the expected size（370 bp）,the homology analysis showed that IL-8 gene was more conservative,which homology was 96.1% to 99.0%.