摘要
目的研究MHCⅡ类反式激活因子(CⅡTA)基因编码区非同义单核苷酸多态性(SNP)位点C19170G(Leu45Val)和C30799G(Ala500Gly)构成的4种不同单倍型eDNA的功能。方法将4种不同单倍型的真核表达载体和空载体分别转染至HeLa细胞。用RT—PCR和间接细胞免疫荧光技术检测未经转染的HeLa细胞、转染4种真核表达载体及空载体的HeLa细胞C1ITAmRNA与3种HLAⅡ类分子(HLA-DR、DP、DQ)的表达,并用流式细胞技术对其表达的3种HLAⅡ类蛋白进行定量分析。结果未经转染和转染空载体的HeLa细胞均无CⅡTA mRNA和3种HLAⅡ类分子的表达,而转染4种不同单倍型真核表达载体的HeLa细胞均出现CⅡTA mRNA表达,并表达3种HLAⅡ类分子。证实了转染4种不同单倍型真核表达载体后的HeLa细胞3种HLAU类分子表达水平差异无统计学意义(P均〉0.05)。结论中国人CⅡTA基因编码区这两个SNP位点的多态性(2个位点氨基酸的改变)不影响CⅡTA反式激活HLAⅡ类基因表达的能力。
Objective To study the function of 4 different haplotypes eDNA which are constructed by two non-homonymy single nueleotide polymorphism (SNP) sites C19170G (Leu45Val) and C30799G (Ala500Gly) in the coding region of human C Ⅱ TA gene. Methods HeLa cells were transfected with eukaryotic expression vectors containing four different haplotypes eDNA. C Ⅱ TA mRNA and HLA class Ⅱ antigen (HLA-DR, DP, DQ) were respectively detected by RT-PCR and indirect cell immunofluoreseence technique in the untransfected and transfected with four eukaryotic expression vectors and empty vectors HeLa cells. The quantity of HLA class Ⅱ antigen were analyzed by flow cytometry. Results No expression of C Ⅱ TA mRNA and HLA class Ⅱ antigen were observed on original HeLa cells and empty vector transfected cells. C Ⅱ TA mRNA expression was emerged, and the expression of HLA class Ⅱ antigen were observed in the HeLa cells transfeeted with eukaryotic expression vectors containing four different haplotypes eDNA. And there were not significantly different with the levels of HLA class Ⅱ antigen expression among HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes eDNA ( P 〉 0.05 ). Conclusion The SNP of Chinese at the sites C19170G(Leu45Val) and C30799G(AlaS00Gly) in the coding site of C Ⅱ TA gene did not influence capability of CⅡTA trans-activating HLA class Ⅱ gene expression.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2009年第6期544-547,共4页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金资助项目(30371283,30872229)