摘要
以人工合成的小鼠免疫球蛋白重链可变区(V_H)基因特异的寡核苷酸片段为引物,用PCR法成功地进行了HIT_3a(抗CD_3单克隆抗体)V_H基因的扩增,并将扩增片段克隆进了pGEM-7Zf(+)质粒DNA中,重组质粒序列测定的结果表明,片段大小为353 bp,基因重排方式为VDJ_4。
By using synthetic oligonucleotide primers specific for V_H gene sequences of mouse IgG, PCR amplification of the V_H gene from the cDNA of HIT3a hybridoma cells, a murine monoclonal antibody specific to CD_3 was successfully performed. The amplified fragment has been cloned into pGEM-7Zf(+)plasmid DNA. Southern analysis shows that the cloned fragment hybridizied strongly to both the universal oligonucleotide and DNA probe of the J region of mouse IgG Hc Analysis of the sequences of recombinant plasmid DNA indicates that the cloned V_H gene is 353 bp in size and VDJ_4 rearrangement in which the D fragment comes from DSP 2.2, and is grouped into Ⅱ B subset.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
1992年第2期131-136,共6页
Acta Academiae Medicinae Sinicae
基金
国家"863"基金
中国医学科学院院校基金部分资助
关键词
免疫球蛋白
PCR
单克隆抗体
V_H gene sequences of IgG
polymerase chain reaction (PCR)
monoclonal antibody
cloning of V_H gene
