人CD38抗原胞外段基因克隆及表达

目的 克隆并表达人CD38抗原分子的胞外估基因。方法 采用RT－PCR法，从高表达CD38抗原的Daudi细胞系中，扩增CD38全长cDNA，并将其插入pGEM-T载体中。重新设计引物，从重组pGEMT载体中，扩增CD38抗体分子的胞外段基因，再亚克隆到表达载体pET28a( )，转化大肠杆菌BL21，用IPTG诱导表达。结果 经酶切鉴定及序列分析表明，克隆的CD38外段基因的序列与文献^1,2]的报道完全一致。将该片段亚克隆到表达载体pET28a( ) ,经IPTG诱导在大肠杆菌BL21中获得表达。结论 获得了人CD38抗原分子胞外段基因及其原核表达产物，对进一步制备单克隆抗体，研究CD38分子的功能具有重要的意义。

Cloning and expression of the extracellular domain of human CD38 antigen gene

Aim To clone and express the 799 bp cDNA fragment of the extracellular domain of human CD38. Methods The cDNA of the human CD38 antigen was amplified from total RNA of Daudi cells by RT PCR, and it was inserted into pGEM T. The cDNA fragment of human CD38 was amplified from the recombinant vector pGEM T, then it was subcloned into expression vector pET28a(+). E. coli BL21 was transfected with recombinant vector and expression was induced with IPTG. Results DNA sequencing showed that cloned 799 bp cDNA sequence was identical with having reported it. The target gene was expressed in E. coli . Conclusion The expressed product of 799 bp cDNA fragment of human CD38 obtained lay the foundation for studing the function of human CD38 antigen and producing anti CD38 monoclonal antibody.