RT-PCR扩增人抗HBsAg抗体Fab段基因  

Amplification of Fab Fragments Genes of Human Antibodies against Hepatitis B Virus by Using RT-PCR

作　　者：唐莉[1] 陈知水[1] 魏来[1] 朱学海[1] 翁国斌[1] 习东[2] 严伟明[2] 陈忠华 

机构地区：[1]华中科技大学同济医学院附属同济医院器官移植研究所,武汉430030 [2]华中科技大学同济医学院附属同济医院临床免疫研究室,武汉430030

出　　处：《华中科技大学学报（医学版）》2004年第2期177-179,共3页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

摘　　要：目的　采用逆转录 聚合酶链反应 (RT PCR)的方法从少量外周血中扩增抗HBsAg抗体重链Fd和κ轻链基因。方法　用RNAex试剂提取细胞总RNA ,以Olig (dT)引导合成cDNA第一链 ,再经PCR扩增的方法 ,以重链Fd和κ轻链基因简并引物 ,直接从人外周血混合淋巴细胞中扩增出抗HBsAg抗体重链Fd和κ轻链基因。 结果　扩增出分子量为 70 0bp的抗HBsAg抗体重链Fd和κ轻链基因。Objective To obtain the anti hepatitis B virus (anti Hbs) antibody Fab genes by using RT PCR. Methods The total cell RNA extracted from peripheral lymphocytes of anti Hbs antibody positive volunteers was transcribed reversely into cDNA. The Fd fragments and light chain genes of human anti Hbs antibody were amplified from cDNA using polymerase chain reaction. Results Fd fragment gene and κchain gene of 700 bp was amplified. Conclusion Fab fragment genes may lay a solid foundation for further research into setting up recombinant adeno associated virus (r AAV).

关 键 词：RT-PCR 基因扩增 抗HBsAg抗体 腺相关病毒载体 外周血 抗体基因 肝移植 

分 类 号：R394.8[医药卫生—医学遗传学]