绵羊ras基因第一、二外显子的扩增及序列测定  

THE AMPLIFICATION AND SEQUENCING OF THE SHEEP ras GENE FIRST AND SECOND EXON

作　　者：刘霄卉[1] 于立新[1] 刘畅[1] 周建华[2] 马学恩[1] 

机构地区：[1]内蒙古农业大学动物科学与医学学院,呼和浩特010018 [2]中国农业科学院哈尔滨兽医研究所,哈尔滨150001

出　　处：《内蒙古农业大学学报（自然科学版）》2011年第4期14-18,共5页Journal of Inner Mongolia Agricultural University(Natural Science Edition)

基　　金：国家自然科学基金项目(30660136)

摘　　要：对JSRV感染羊及健康绵羊H-ras、N-ras、K-ras基因第一、二外显子区克隆并测序,研究其变异状况。方法:本研究从JSRV感染羊及健康绵羊肺组织中提取基因组DNA,应用PCR技术扩增Hras、Nras、Kras基因第一、二外显子序列,并通过T-A克隆的方法将其克隆入pMD-18T载体中,鉴定为阳性的重组质粒进行序列测定分析。结果:与其他物种ras基因相比,所克隆的ras基因与其他物种之间的差异率较小,基本保守,且与牛的ras基因同源性最高。所克隆的H-ras、K-ras基因在第一、二外显子区域序列完全相同;N-ras第81位氨基酸A-V的不同;但试验中检测的2例JSRV感染羊与健康绵羊的ras无论是在核酸水平上,还是在氨基酸水平上均未发现突变。结论:克隆测序了绵羊ras基因第一、二外显子,为下一步实验奠定了基础。Objective：To clone and sequenceJSRV infection in sheep and healthy sheep Hras,Nras,Kras gene first and second exon and to study the mutation of the gene.Methods：In this study,the H-ras,N-ras,K-ras gene first and second exon DNA was extracted from healthy sheep and sheep infected with JSRV lung tissue and amplified with PCR.The PCR product was cloned into pMD-18T vector plasmid.Than the recombinant plasmid was identified and sequenced.Results：Compared with other species,ras gene had little difference among species and highly homology with the cattle ras gene.The H-ras,K-ras gene cloned was identical in the first and second exon region sequence,while the N-ras had a A-V difference in the 81th amino acid.There were no mutation ras gene between the two cases of JSRV infection in sheep and healthy sheep neither in nucleic acid level nor in the amino acid level.Conclusion： The cloning and sequencing of the first and second exon of the sheep ras gene laid a good foundation for further study.

关 键 词：RAS基因 第一、二外显子 克隆 序列分析 

分 类 号：S852.3[农业科学—基础兽医学]