| 两种抗α毒素单链抗体基因的序列分析 |
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| 引用本文： | 赵宝华,许崇波.两种抗α毒素单链抗体基因的序列分析[J].河北师范大学学报(自然科学版),2001,25(1):109-112. |
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| 作者姓名： | 赵宝华 许崇波 |
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| 作者单位： | 1. 河北师范大学生命科学学院,
2. 解放军军需大学军事兽医研究所, |
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| 基金项目： | 全军医药卫生科学基金课题!(98Q0 76 ) |
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| 摘 要： | 应用RT－PCR技术，从两株分泌具有中和活性的抗A型产气荚膜酸菌α毒素单克隆抗体（McAb)的杂交瘤细胞株2E3和1A8中，分别扩增出抗体VH和VL基因，用Linker(Gly4Ser）3基因，将VH和VL基因连接成ScFv基因2E3－ScFv和1A8－ScFv，并将其克隆至pGEM-T载体中，经核苷酸序列分析证实，VH和VL基因以及Linker基因拼接正确，2E3－ScFv基因全长为729bp，经计算机分析，VH和VL基因均为新发现的基因序列，符合功能性重排的鼠抗体可燮区基因特征，2E3－ScFv的VH和VL基因分别属于鼠免疫球蛋白重链Ⅱ（B）和轻链kⅢ家簇；而1A8－ScFv的VH和VL基因分别属于鼠免疫球蛋白重链Ⅱ（A）和轻链кⅥ家簇。
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| 关 键 词： | α毒素 单链抗体 基因克隆 序列分析 产气荚膜梭菌 单克隆抗体 RT－PCR技术 |
| 文章编号： | 1000-5854(2001)01-0109-04 |
| 修稿时间： | 2000年4月5日 |
Sequence Analysis of ScFv Gene for Anti-alpha-toxin |
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| ZHAO Bao-hua,XU Chong-bo.Sequence Analysis of ScFv Gene for Anti-alpha-toxin[J].Journal of Hebei Normal University,2001,25(1):109-112. |
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| Authors: | ZHAO Bao-hua XU Chong-bo |
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| Abstract: | The VH and VL genes were amplified from two hybridoma cell lines producing mouse McAb against alpha-txoin of Clostridium perfringens type A by RT-PCR. The VH and VL genes were connected through a flexible Linker (Gly4Ser)3,and the VH-Linker-VL(ScFv) fusion gene was cloned into a clone vector pGEM-T. The 2E3-ScFv and 1A8-ScFv gene were sequenced and analyzed by computer. The ScFv genes consist of 726 bp and 729 bp respectively.Both VH and VL genes were functionally rearranged mouse immunoglobulin variable region genes and appeared to be new genes. According to Kabat classed method,the VH gene segment and VL gene segment of 2E3-ScFv belong to the mouse Ig heavy chain subgroup Ⅱ (B) and к chain subgroup Ⅲ respectively,but the VH gene segment and VL gene segment of 1A8 -ScFv belong to the mouse Ig heavy chain subgroup Ⅱ (A) and к chain subgroup Ⅵ respectively. |
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| Keywords: | alpha toxin ScFv gene cloning sequence analysis |
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