鸡脾细胞总RNA的提取及其V_H、V_L基因的扩增

Extraction of Total RNA from Chicken Splenic Cell and Amplification of V_H and V_L Gene Fragments

由鸡脾脏淋巴细胞中提取总RNA,反围录合成cDNA第一链,以cDNA为模板,根据鸡抗体V_H、V_L基因中的5’端和J端序列,设计并合成体外扩增V_H、V_L基因的两对引物,进行RT-PCR,成功地扩增出V_H、V_L基因片段。为鸡源ScFv噬菌体库的构建打下基础。

Total RNA were extractec from chickn slpenic lymphocyte and cDNA were synthesiaed by reverse transcription. using two sets of oligonucleotide primer which were designed and synthesized according to 5' and J region gene sequences of chicken immunoglobulin variable region, we amplified VH and VL gene repertoire successfully with cDNAs' templates by RT-PCR. It will be useful for constructing phage display library of chicken repertoire ScFv.