摘要
目的：克隆黑色素瘤特异性单克隆抗体ＨＢ８７６０可变区基因．方法：从培养的可分泌人黑色素瘤特异性单克隆抗体的杂交瘤细胞ＨＢ８７６０提取总ＲＮＡ，反转录成ｃＤＮＡ，用合成的一对寡核苷酸引物从中扩增了抗体可变区基因，并克隆入ｐＵＣ１９，挑选出阳性克隆后进行了核苷酸序列分析．结果：所克隆基因分别长３６０ｂｐ和３３０ｂｐ，编码１２０个和１１０个氨基酸，含三个ＣＤＲ区和四个ＦＲ区，并含有维持抗体结构所必需的两个半胱氨酸．计算机分析与已发表的小鼠抗体可变区基因有较高同源性．结论：所克隆基因可编码小鼠抗体可变区．
Objective: Cloning of the variable region genes of a human melanoma specific monoclonal antibody. Methods: Total RNA was isolated from a cultured hybridoma cell line HB8760 which secretes human melanoma specific monoclonal antibody, and followed by reverse transcription into cDNA. Then PCR was used to amplify the immunoglobulin variable region genes of the antibody. The PCR product was cloned into pUC19 plasmid and sequenced. Results: The cloned genes consist of 360 bp and 330 bp respectively and encode 120 and 110 amino acids. There are three CDRs, four FRs and the charicteristic cystein residues within the genes. The genes are homologues to the mouse Ig genes published in EMBL gene bank 1995. Conclusions: The data indicate that the cloned genes encode the variable region of the mouse antibody.
出处
《第四军医大学学报》
1997年第3期225-227,共3页
Journal of the Fourth Military Medical University
