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RT-PCR扩增人源性抗腮腺炎病毒抗体轻链和重链Fd基因
RT-PCR Amplification of Light Chain and Heavy Chain Fd Fragment of Genes from Human Antibodies Against Mumps Viruses
【摘要】 目的 克隆人源性抗腮腺炎病毒抗体轻链和重链Fd基因。方法 从腮腺炎患者和腮腺炎抗体IgG阳性者中各抽取其外周血 3ml,分离出淋巴细胞后 ,提取总RNA ,逆转录成cDNA。设计人抗体轻链κ、λ和重链Fd基因的引物进行聚合酶链反应 (PCR) ,扩增轻链和重链Fd段基因。结果 从 4 8名腮腺炎病毒IgG抗体阳性者中 ,共提取高质量RNA约 1 1 0 μg,经RT PCR分别扩增出约 70 0bp大小的κ、λ和Fd基因。结论 从人淋巴细胞中成功克隆出抗体轻链κ、λ和重链Fd基因 ,为制备抗体Fab重组表达载体 ,进一步构建抗腮腺炎病毒噬菌体抗体库打下基础
【Abstract】 Objective To clone gene of light chain and heavy chain Fd fragment from human antibodies against mumps viruses. Methods Extracted total RNA from peripheral blood lymphocytes (PBLs) of mumps patients and health population with positive mumps IgG, and reverse transcribed into cDNA. Gene of light chain and Fd fragment genes of heavy chain of the immunoglobulin were amplified by PCR using primers for κ,λ and Fd. Results A total of 110 μg total RNA of high quality was extracted from PBLs and. about 700 bp κ,λ and Fd genes were respectively obtained. Conclusion Successful colonizing of light chain κ,λand heavy chain Fd genes will provide a foundation for further construction of Fab recombinant expression vector.
【Key words】 Mumps virus; Reverse transcriptase polymerase chair reaction; Genes;
- 【文献出处】 华南预防医学 ,Guangdong Journal of Health and Epidemic Prevention , 编辑部邮箱 ,2003年06期
- 【分类号】R392.1
- 【下载频次】63