应用PCR从丙型肝炎患者外周血B淋巴细胞扩增抗体基因-东南大学学报医学版

应用PCR从丙型肝炎患者外周血B淋巴细胞扩增抗体基因

单位：1.南京铁道医学院,微生物学与免疫学教研室,江苏,南京,210009； 2.南京铁道医学院,生物学教研室,江苏,南京,210009

分类号：R512.6+3, R392.12, R331.1+44

出版年·卷·期（页码）：2000·19·第一期（17-20）

摘要：

用常规RTPCR法,以1组人抗体重链和轻链简并引物,直接从5名丙型肝炎患者外周血混合淋巴细胞中扩增出抗体重链Fd基因和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,为构建噬菌体抗体库奠定了基础.

Employing routine RT?PCR,the genes of human immunoglobulin Fd and κ light chains were amplified from hepatitis C patients peripheral blood lymphocytes by sets of oligonucleotide primers. 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 satisfactory. The high purification of the total RNA of the cells is not necessary to RT?PCR.

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