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从携带人转铁蛋白单链抗体基因载体转化的大肠杆菌菌株中提取质粒,经电泳分析及筛选、纯化出线状质粒DNA,定量后以载体上人转铁蛋白单链抗体基因插入子两边的已知DNA序列经合成后作为引物进行测序扩增凝胶电泳,将得到的人转铁蛋白单链杭体基因序列重轻链可变区部分同基因库中鼠抗体重轻链进行同源比对,证明人转铁蛋白单链抗体基因确由鼠抗体重轻链可变区基因构成,在此基础上,进一步对基因核苷酸序列的可变区结构特征作推断分析,并推导出其对应的氨基酸序列.
Plasmids were extracted from Escherichia coli strains carrying human transferrin single-chain antibody gene vectors and purified by electrophoresis and screening. Linear plasmid DNAs were quantified and quantified as either the human transferrin single-chain antibody gene insert The DNA sequence was synthesized and sequenced as a primer to amplify the gel electrophoresis, and the resulting heavy chain variable region of the heavy chain of the human transferrin single-stranded gene was homologous aligned with the heavy chain of the mouse antibody in the gene bank , It was proved that the human transferrin single-chain antibody gene was composed of the murine antibody heavy and light chain variable region genes. On the basis of this, the structural features of the variable region of the nucleotide sequence of the gene were further inferred and deduced Of the amino acid sequence.