PCR扩增人外周血淋巴细胞抗体基因的探讨  

Studies on Conditions about Amplification of Immunoglobulin Genes from Human Peripheral Blood Lymphocytes by PCR

作　　者：张建琼[1] 单祥年[1] 张雪萍[1] 林陵[1] 谢维[1] 

机构地区：[1]南京铁道医学院微生物免疫学教研室

出　　处：《南京师大学报（自然科学版）》1999年第1期73-76,共4页Journal of Nanjing Normal University(Natural Science Edition)

基　　金：江苏省科技发展基金

摘　　要：采用ＲＴ－ＰＣＲ法，以一组人抗体重链和轻链简并引物，直接从人外周血混合淋巴细胞中扩增出抗体重链Ｆｄ和κ轻链基因．结果提示：用Ｔｒｉｚｏｌ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，是值得优先采用的方法．细胞总ＲＮＡ的提取不必追求高纯度，少量外周血即足以满足构建抗体库所需，简并引物可获得良好的扩增效果。Employing routine RT-PCR ,the genes of human immunoglobulin Fd and κ light chains were amplified from human peripheral blood lymphocytes by sets of oligonucleotide primers designed by kang.These results suggest that by using Trizol reagant isolated the total RNA of the cells,using Olig(dT) primer reverse transcription synthesized the cDNA first-strand were good methods.The total RNA of the cells is not necessary high purification for RT-PCR.

关 键 词：聚合酶链反应 抗体基因 外周血 淋巴细胞 PCR 

分 类 号：R392.11[医药卫生—免疫学]