| 摘要: |
| 从蝎毒腺中提取总RNA，通过RT－PCR得到东亚钳蝎神经毒素BmKCT的cDNA，将其连接取到pUCm-T载体中。序列分析表明，该基因开放阅读框架编码59个氨基酸残基，其中24个为信号肽，其余35个为成熟肽，与Genebank上登录的相同。将蝎神经毒素cDNA再经PCR扩增除去除信号肽序列，克隆到pTrcHisA表达载体中，转化大肠杆菌BL21（DE3），经IPTG诱导可高效表达分子质量为10 ku左右的融合蛋白。表达产物约占菌体总蛋白的25％。 |
| 关键词: 蝎毒素 cDNA 融合表达 |
| DOI： |
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| 基金项目:河南省重点科技攻关项目(0223032300) |
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| Cloning and expression of a neurotoxin from chinese scorpion Buthus martensii Karsch in Escherichia coli |
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| Abstract: |
| A neurotoxin cDNA was cloned from Chinese scorpion Buthus martensii Karsch by RT-PCR.The cDNA was cloned into the pUCm-T vector and sequenced.It has a ORF encoding 59 amino acid residues and a 24 residues signal peptide.The cDNA encoding the mature peptide was amplified by PCR and was cloned into pTrcHisA vector.The recombinant vector was transformed into E.coli BL21(DE3).The E.coli highly expressed the fusion protein whose mollecular weight is 10 ku,after induced by 0.1 mol/L IPTG.The expressed protein was accumulated up to more than 25% of total bacterial protein. |
| Key words: scorpion toxin cDNA fusion expression |
