从人外周血B淋巴细胞中应用PCR扩增人抗体基因

本文用常规ＰＣＲ法和半套式ＰＣＲ法，以一组人抗体重链和轻链引物，直接从人外周血淋巴细胞中扩增出抗体重链Ｆｄ基因和轻链基因。一些常规ＰＣＲ法不能直接扩增的人抗体基因，用半套式ＰＣＲ法扩增却得到了阳性结果。扩增的抗体基因的分子量与国内外同类报道一致。本文结果提示，在建立抗体基因文库时，半套式ＰＣＲ法能进一步丰富扩增的抗体基因的多样性

Amplification of Immunoglobulin Genes from Human Peripheral Blood Lymphocytes by PCR

Employing routine PCR and half nested PCR, the genes of human immunoglobulin Fd and light chains were amplified from human peripheral blood lymphocytes by sets of oligonucleotide primers designed by Kang. Some primers could not directly amplify antibody genes from cDNA templates. So, a hald nested PCR method was used with primers designed to maximize homology with human immunoglobulin signal peptide sequences as the first round PCR. The molecular weight of products amplified by routine and half nested PCR is identical to that reported by other scholars. These results suggest that half nested PCR might be helpful for maximizing the diversity of heavy and light chain genes of human immunoglobulin.