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 。
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.
出处
《华南预防医学》
2003年第6期10-12,共3页
South China Journal of Preventive Medicine
基金
2 0 0 2年广东省医学科研基金 (A2 0 0 2 666)
深圳市科技计划基金项目
关键词
腮腺炎病毒
逆转录聚合酶链反应
基因
Mumps virus
Reverse transcriptase polymerase chair reaction
Genes
