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目的构建人 CD15 4基因真核细胞表达载体。方法采用 RT- PCR方法 ,从激活的人外周血淋巴细胞的总c DNA中扩增得到 82 0 bp的人 CD15 4c DNA片段 ,再用 Bam H 和 Eco R 双酶切后定向克隆到真核细胞表达载体pc DNA3.1中 ,限制性内切酶酶切分析重组质粒和 DNA序列分析。结果人 CD15 4c DNA已经正确克隆到真核细胞表达载体pc DNA3.1中。结论本研究为探讨 CD15 4分子与淋巴细胞活化的关系及其信号传导机制 ,为进一步用于抗移植排斥反应的研究 ,或对由于 CD15 4遗传缺陷所致的免疫缺陷病的基因治疗奠定了基础
Objective To construct eukaryotic expression vector of human CD15 4 gene. Methods A total of 82 0 bp human CD15 4c DNA fragments were amplified by RT-PCR from total c DNA of activated human peripheral blood lymphocytes. After digested with Bam H and Eco R, the recombinant plasmid was cloned into eukaryotic cells Expression vector pcDNA3.1, restriction endonuclease digestion analysis of recombinant plasmids and DNA sequence analysis. Results The human CD15 4c DNA was correctly cloned into the eukaryotic expression vector pc DNA3.1. Conclusion This study was designed to investigate the relationship between CD15 4 and lymphocyte activation and its signal transduction mechanism and to lay foundation for the further study of anti-graft rejection or gene therapy of immunodeficiency caused by CD15 4 genetic defect