摘要
目的:扩增TLR7主要编码区外显子3(exon 3)的全长基因片段并进行基因多态性分析,筛选健康人群中TLR7基因的主要SNP位点。方法:采用酚-氯仿抽提方法从28例健康女性血样中提取基因组DNA,采用长片段扩增方法分别扩增TLR7 exon 3区的3个片段,测序、拼接后进行多态性分析。结果:采用LA-Taq酶体系成功扩增TLR7 exon 3区基因片段;经与Genbank数据库中TLR7参考序列比较,我国女性TLR7基因序列高度保守,仅出现了4个点突变,并发现1个SNP位点RS3853839,表现为GG、CG和CC三种基因型。结论:建立了TLR7编码区基因扩增方法,筛选到1个TLR7 SNP位点RS3853839,可为分析TLR7多态性与多种病毒感染性疾病的关系提供参考。
Objective： To amplify the coding region of Toll-like receptor 7 （TLR7）, investigate the polymorphisms and screen major single nucleotide polymorphism （SNP） of TLR7 gene in Chinese females. Methods： Genomic DNA was obtained from 500 ul blood by phenol-chloroform extraction. Three long fragments of TLR7 gene were amplified with LA-Taq polymerase and special primers. PCR products were sequenced and assembled. Results： The full-length of TLR7 exon-3 region was successful amplified by LA-Tag polymerase. The TLR7 sequences of Chinese female were very conservative. Only four mutations and one SNP RS3853839 were detected in 28 females. There were three genotypes GG, CG and CC in RS3853839. Conclusion： The amplification method of TLR7 coding region gene was established, the SNP RS3853839 was found in health females, and it may provide reference for study on TLR7 polymorphism in virus diseases and immunity diseases.
出处
《现代生物医学进展》
CAS
2013年第6期1055-1058,共4页
Progress in Modern Biomedicine
基金
陕西省科技攻关项目(2009K17-05)
关键词
TLR7
扩增
基因
多态性
Toll-like receptor 7
PCR
Gene
Single nucleotide polymorphism
