牛白介素10的克隆、测序及其在CHO细胞的表达  

Cloning,Sequencing and Eukaryotic Expression of Bovine Interleukin-10 Gene in CHO Cells

作　　者：敖敦格日勒[1] 温德宝 百溪英一 格日勒图[1] 

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]阿巴嘎旗动物卫生监督所,内蒙古阿巴嘎旗011400 [3]日本农业技术研究机构动物卫生研究所免疫研究部

出　　处：《动物医学进展》2014年第4期6-9,共4页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31360602);内蒙古自然科学基金项目(2011BS0408)

摘　　要：用RT-PCR方法从健康牛外周血淋巴细胞中扩增出牛白介素10(bIL-10)基因并进行测序,发现其核苷酸序列全长为537bp,编码179氨基酸,具有很好的遗传保守性。将该基因克隆到表达载体pcDNA3.1/V5-His Vectors A中,构建真核表达重组质粒pcDNA3.1/V5-His A/IL-10,将该质粒转染到CHO细胞中,采用SDS-PAGE和Western blot方法检测培养物上清和沉淀物中bIL-10的表达,结果表达的bIL-10大小约为20ku。本试验为bIL-10生物学特性及其基因多态性的研究奠定了基础。Bovine IL-10 （bIL-10） gene was amplified by RT-PCR from peripheral blood lymphocytes of healthy bovine in this study. The sequencing results indicated that the bovine IL-10 gene was 537 bp in full length, and translated by 179 amino acids, with a very stable genetic conservation. This gene was inserted into the eukaryotic expression vector pcDNA3/VS-His Vectors A, and successfully constructed the eukaryotie expression vector of peDNA/V5-bIL-10, and then transfected into the CHO cells for expression. Then expression products in culture of supernatant and precipitate were identified by SDS-PAGE and Western blot methods, which had a molecular weight of approximately 20 ku. These experiments lay a foundation for further study on the biological characteristics and gene polymorphism of the bIL-10.

关 键 词：牛白介素-10 克隆 测序 真核表达 

分 类 号：Q789[生物学—分子生物学]