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利用 RT-PCR 和噬菌体表面展示技术,直接从乙肝病毒核心抗体(抗 HBc)阳性患者淋巴细胞中提取总 RNA,反转录成 cDNA;合成全套人抗体可变区引物扩增抗体可变区基因,并将重、轻链可变区基因进行拼接装配成单链抗体(ScFv)基因,重组于噬菌粒载体 pHEN1,转化抑制型大肠杆菌 E.coliTG1,以辅助噬菌体援救后,构建成人源性单链噬菌体库。库容量达106。
Total RNA was directly extracted from lymphocytes of hepatitis B virus-positive (anti-HBc) -positive patients by RT-PCR and phage display technology and then transcribed into cDNA. A complete set of human antibody variable region primers were synthesized to amplify the antibody variable region genes , And the heavy and light chain variable region genes were spliced and assembled into the single-chain antibody (ScFv) gene, recombined in the phagemid vector pHEN1 and transformed into E. coli E.coli TG1 to aid human phage rescue to construct human-derived Single-stranded phage libraries. Library capacity of 106.