摘要
目的 :构建g3pN1 hCD2 0跨膜及胞外区基因表达载体 ,并在大肠杆菌中进行高效融合表达。方法 :利用反转录PCR和PCR方法 ,分别从Daudi细胞和M13K0 7噬菌体中扩增hCD2 0基因和编码g3pN1蛋白N1端的基因 ,并重组到pTIG Trx表达载体中 ,在大肠杆菌中融合表达。结果 :表达产物可被抗CD2 0分子的单克隆抗体 (mAb)识别。结论 :成功地表达并鉴定了目的蛋白。
AIM: To construct the expression vector containing transmembrane domain gene of human CD20 and g3pN1 gene and express the fusion gene high-efficiently in E.coli. METHODS: The human CD20 gene and g3pN1 domain gene were amplified by RT-PCR and PCR from Daudi cells and M13K07 phage antibody library, respectively, and then cloned into expression vector pTIG-Trx.The constructed expression vector was expressed in E.coli. RESULTS: Western blot analysis showed that expressed product could bind to anti-CD20 mAb. CONCLUSION: The pTIG-GS has been constructed and expressed successfully in E.coli, which lays the foundation for further screening anti-CD20 antibody from phage antibody library.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2004年第4期481-483,共3页
Chinese Journal of Cellular and Molecular Immunology
