摘要
目的 应用TA克隆技术构建含HLA-DRα基因的载体,作为实时荧光定量反转录聚合酶链反应(RT-PCR)法检测 HLA-DRα基因的标准模板。方法 利用RT-FCR法,从提取的外周血单核细胞总RNA中特异地扩增含635bp的HLA- DRαcDNA片段,并将其克隆到TA载体。结果 用限制性内切酶法和测序方法证实扩增片段为目的片段。结论 成功地构建了 HLA-DRα基因片段TA克隆载体。为实时荧光定量RT-PCR法检测HLA-DRαmRNA表达提供标准模板。
Objective： To construct of HLA-DRα gene vector by TA cloning technique, and the vectoris acted as the standard templat detected HLA-DRα gene with a real-time quantitative RT-PCR method. Methods： The 635bp region of HLA-DRα cDNA was amplified by RT-PCR from extracted monocytes total RNA. The PCR product was cloned into TA cloning vector. Results： The TA cloning vector was identified being HLA-DR, fragment, with the restriction enzyme and sequencing methods. Conclusion： The HLA-DR.gene TA cloning vector was contracted successfully.
出处
《湖北中医学院学报》
2005年第4期31-32,共2页
Journal of Hubei College of Traditional Chinese Medicine
