期刊论文

中文

中国农学通报

1000-6850

2010

26

23

22-25

国家自然科学基金“应用鼠源性高亲和力噬菌体Fab抗体筛选和克隆新的血清学H-Y抗原基因”（30901090/C120209）； 湖南省教育厅科学研究优秀青年项目“利用高亲和力噬菌体抗体筛选血清学H-Y抗原”（09B046）；

本校为第一机构

为了扩增多样性Fab抗体基因产物,保证噬菌体抗体库的足够库容。分离C57BL/6鼠的脾脏,并制备脾细胞悬液,提取其总RNA,逆转录为cDNA,利用普通PCR和Touch down（TD）PCR扩增鼠Fab基因产物,通过琼脂糖凝胶电泳验证PCR产物,比较分析2种方法的扩增效果。结果显示:轻链和重链Fd基因的PCR产物大小约为680bp,与理论值相符。电泳结果显示,TD-PCR获得8个重链Fd基因和6个轻链基因条带,普通PCR获得7个重链Fd基因和5个轻链基因条带。在普通PCR产物中,轻链基因引物VkB未能扩增出相应基因,而TD-PCR扩增出该基因。TD-PCR扩增出引物ⅢA和ⅢC的对应基因,而普通PCR却未见对应结果,普通PCR扩增出ⅢB引物的对应基因,TD...

To amplify various products of Fab antibody genes with diversity,guarantee enough reservoir capacity of phage antibody library.The spleen of C57BL/6 mouse was isolated,spleen cell suspension was prepared,total RNA was extracted,cDNA synthesis was accomplished by reverse transcription.The product of Fab antibody was amplified by PCR and Touch down PCR,the products were analyzed by agarose gel electrophoresis,the author performed a comparative analysis of two PCR methods.the products of light chain and high chain gene was about 680 bp,and the result corresponded with the theoretical value.The re...