| 摘要: |
| 通过RT-RCR得到线虫fat-1基因全长，将其N端亲水区克隆至原核表达载体pGEX-4T-2中，构建重组质粒pGEX-fat1-N，将其转化大肠杆菌，并进行诱导表达。对表达产物GST-fat1-N融合蛋白进行纯化，制备其抗体，并对抗体效价进行了检测。结果表明，线虫fat1基因能在大肠杆菌中表达，制备的抗体能识别在原核表达系统内表达的GST-fat1-N融合蛋白，且效价很高，达到1∶107。 |
| 关键词: fat-1基因 线虫 原核表达 抗体制备 |
| DOI： |
| 分类号: |
| 基金项目:中国科学院知识创新工程青年科学家专项基金 |
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| Prokaryotic expression of the C.elegans fat-1 gene and preparation of antibody |
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| Abstract: |
| Caenorhabditis elegans fat-1 gene was obtained by RT-PCR,and the sequences of fat-1 gene N terminal were cloned into fusion expression vector pGEX-4T-2 to generate recombinant plasmid pGEX-fat1-N.After expressing in Escherichia coli,the expressed product GST-fat1-N fusion protein was purified and used to immunize rabbit.The result showed C.elegans fat1 gene could be expressed in E.coli,and the antibody against GST-fat1-N protein had a good affinity and selectivity by enzyme-linked immunosorbent assay. |
| Key words: fat-1 gene Caenorhabditis elegans prokaryotic expression antibody preparation |
