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目的:克隆人T-bet基因全长编码区的cDNA,并进行鉴定和测序分析,为从转录水平研究Th1/Th2平衡及其与免疫相关性疾病发生发展的关系奠定基础。方法:采用RT-PCR方法,从人外周血淋巴细胞获得T-bet基因的cDNA;连接至pGEM-Teasy载体,导入大肠杆菌DH5α并选择阳性克隆;应用M13F/R通用引物进行双向反应测序,以作序列鉴定。结果:扩增得到的T-bet基因cDNA全长1 670 bp,包括编码区1 607 bp,编码530个氨基酸残基,与Gen-bank中发表的序列完全一致。结论:获得了人T-bet基因的克隆,为进一步研究其生物学功能构建了基础平台,为寻找多种免疫性疾病的有效治疗提供新思路。
OBJECTIVE: To clone cDNA of full-length coding region of human T-bet gene and identify and sequence it for the study of the relationship between Th1 / Th2 balance and the development of immune-related diseases at the transcriptional level. Methods: cDNA of T-bet gene was obtained from human peripheral blood lymphocytes by RT-PCR, ligated to pGEM-Teasy vector, introduced into E. coli DH5α and selected positive clones. The M13F / R universal primer was used to perform bidirectional reaction sequencing. For sequence identification. Results: The full-length cDNA of T-bet gene was 1 670 bp in length, including 1 607 bp in coding region encoding 530 amino acid residues, which was completely identical with the published sequence in Gen-bank. Conclusion: The cloning of human T-bet gene was obtained, which provided a basic platform for further study of its biological functions and provided new ideas for the effective treatment of various immune diseases.