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目的 :克隆人CD2 0胞外区基因 (cdw)和丝状噬菌体 (M13K0 7)G3蛋白N端结构域 (pⅢN1)基因 ,并在大肠杆菌中进行高效融合表达。方法 :利用逆转录PCR和PCR方法分别克隆CD2 0胞外区基因和pⅢN1基因 ,然后将二者融合克隆入pTIG Trx表达载体 ,在大肠杆菌中进行可溶性表达。结果 :可溶性表达产物占细菌可溶性蛋白的约2 5 % ,表达产物可被抗CD2 0分子的单克隆抗体识别。结论 :成功地表达并鉴定了人CD2 0胞外区蛋白 ,为利用噬菌体抗体库进行抗CD2 0抗体的筛选奠定了基础。
OBJECTIVE: To clone the gene encoding the N-terminal domain (pⅢN1) of human CD20 extracellular region (cdw) and filamentous bacteriophage (M13K0 7) G3 and express it in E. coli. Methods: CD2 o extracellular region gene and pⅢN1 gene were cloned by reverse transcription PCR and PCR respectively. The two fusion proteins were cloned into pTIG Trx expression vector and expressed in E. coli. Results: The soluble expression product accounted for about 25% of the bacterial soluble protein, and the expression product could be recognized by the anti-CD20 molecule monoclonal antibody. Conclusion: The human extracellular domain of CD20 was successfully expressed and identified, which laid the foundation for the screening of anti-CD20 antibody using the phage antibody library.