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目的 分离日本血吸虫单克隆抗独特型抗体 NP30重链可变区 (VH)基因并测定其序列。方法 根据鼠免疫球蛋白重链可变区基因 FR1和 FR4序列的保守性 ,化学合成体外扩增 Ig重链可变区基因的数对引物。以日本血吸虫单克隆抗独特型抗体 NP30的杂交瘤细胞株基因组 DNA为模板 ,扩增 VH 基因 ,将其克隆入 p U C19载体 ,重组子用 Sanger’s双脱氧链终止法测定序列 ,将序列与Gene Bank中已发表的抗体序列比较。结果 VH 基因全长 35 7bp,属鼠免疫球蛋白重链第 亚类 ,由种系基因 V、Dsp2 .8与 JH4重排而来。该 VH 基因序列已被 Gene Bank收录 (accession NoAF2 82 6 2 2 )。结论 该 VH 基因为日本血吸虫单克隆抗独特型抗体 NP30重链可变区基因
Objective To isolate the VH30 gene of the monoclonal anti-idiotypic antibody NP30 of Schistosoma japonicum and determine its sequence. Methods According to the conservation of the FR1 and FR4 sequences in the variable region of murine immunoglobulin heavy chain, several pairs of primers were designed to amplify Ig heavy chain variable region genes in vitro. The VH gene was amplified from the genomic DNA of the hybridoma cell line of Schistosoma japonicum monoclonal anti-idiotypic antibody NP30 and cloned into pUC19 vector. The recombinant was sequenced by Sanger’s dideoxy chain termination method. Published antibody sequences in Bank. Results The VH gene was 35 7 bp in length, belonging to the murine immunoglobulin heavy chain subgroup and rearranged by the germline genes V, Dsp2. 8 and JH4. This VH gene sequence has been included in Gene Bank (accession NoAF2 82 6 2 2). Conclusion The VH gene is the gene of NP30 heavy chain variable region of the monoclonal anti-idiotypic antibody of Schistosoma japonicum