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目的构建含有HLA-DRA和DRB4*01030101基因全长开放阅读框的cDNA克隆载体。方法用RT-PCR方法从人外周血白细胞中获得HLA-DRA和DRB4*01030101的全长编码基因,分别克隆至pMD18-T载体,并进行序列测定和比对分析。结果从人外周血白细胞中扩增出HLA-DRA和DRB4*01030101含全长开放阅读框的cDNA,分别克隆至pMD18-T载体,经序列测定和比对分析证实,HLA-DRA和DRB4*01030101编码基因与文献报道序列完全一致,阅读框与设计相符。结论成功构建了含有HLA-DRA和DRB4*01030101基因全长开放阅读框的cDNA克隆载体,为体外表达HLA-DRB4抗原分子及研究其分子间相互作用奠定了实验基础。
Objective To construct a cDNA cloning vector containing the full-length open reading frame of HLA-DRA and DRB4 * 01030101 genes. Methods The full-length coding genes of HLA-DRA and DRB4 * 01030101 were obtained from human peripheral blood leucocytes by RT-PCR and cloned into pMD18-T vector respectively for sequence analysis and comparison analysis. Results The cDNAs containing the full-length open reading frame of HLA-DRA and DRB4 * 01030101 were amplified from human peripheral blood leukocytes and cloned into pMD18-T vector respectively. The sequence analysis and comparison confirmed that HLA-DRA and DRB4 * 01030101 The coding gene is identical to the reported sequence in the literature, and the reading frame is in accordance with the design. Conclusion The cDNA cloning vector containing the full-length open reading frame of HLA-DRA and DRB4 * 01030101 gene was successfully constructed, which laid the experimental foundation for the in vitro expression of HLA-DRB4 antigen molecules and their intermolecular interactions.