摘要
采用反转录PCR和PCR方法分别克隆P185HER2n/eu胞外区基因和噬菌体M13KO7g3p-N1结构域基因,然后将二者偶联入pET-22b(+)载体中,在大肠杆菌中进行融合表达。可溶性目的蛋白表达量占细菌可溶性表达产物总量的30%左右,并通过镍亲和层析纯化出目的蛋白。以上结果为从噬菌体抗体库中筛选抗P185HER2/neu的抗体奠定了基础。
The extracellular region gene of human P185HER2/neu and the gene of g3p-N1 domain were amplified by RT-PCR and PCR from sk-br-3 cell and M13KO7 phage respectively. Fused to 3'-terminus of the gene of g3p-N1 domain, the extracellular region gene of human P185HER2/neu was high-efficiently expressed in host E.coli by vector pET-22b(+). The fusion protein attaining to the 30% of the total protein which could be soluble expressed in E.coli was conveniently purified by chelating column (HisTrap HP Columns). This research made the basis of screening anti-P185HER2/neu antibody from recombinant phage-display antibody library.
出处
《生物技术通讯》
CAS
2005年第3期252-254,共3页
Letters in Biotechnology
