[Gene cloning, heterologous expression and activity identification of latroeggtoxin-Ⅵ].
[Gene cloning, heterologous expression and activity identification of latroeggtoxin-Ⅵ].
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DOI：
10.13345/j.cjb.200245
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发表时间：
2021-02-25
期刊：
影响因子：
--
通讯作者：
Wang, Xianchun
中科院分区：
文献类型：
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作者：
Yan, Shuai;Tang, Xiaochao;Wang, Xianchun
间斑寇蛛的一个显著特点是其毒性成分不仅存在于毒腺中，而且存在于毒腺外的组织中，甚至存在于卵中。对蜘蛛毒腺外毒素的研究可以加深对蜘蛛毒素的认识，发现新的具有重要应用前景的先导分子。为了探索L.利用生物信息学方法，从L. tredimguttatus卵的转录组中挖掘出编码肽毒素的基因序列。采用3 '-RACE结合巢式PCR的策略成功地异源表达了该基因。生物学活性分析表明，表达的肽毒素（latroeggtoxin-Ⅵ，LETX-Ⅵ）能抑制ND 7/23细胞Na+通道电流，促进PC 12细胞多巴胺释放，对美洲大蠊、金黄色葡萄球菌和白色念珠菌等细菌和真菌无明显毒性。说明LETX-Ⅵ是一种哺乳动物特异性神经毒素，在开发神经生物学研究的工具试剂和治疗相关疾病的药物方面具有潜在的应用前景。
One of the distinct characters of Latrodectus tredecimguttatus is that its toxic components exist not only in the venomous glands, but also in the tissues outside the venomous glands and even in the eggs. Investigation on the toxins outside the venomous glands can deepen our understanding of spider toxins and discover new lead molecules with important application prospects. In order to explore the low-abundance proteinaceous toxins in the L. tredecimguttatus eggs, we used bioinformatic strategies to mine a gene sequence encoding a peptide toxin from the transcriptome of L. tredecimguttatus eggs, and then heterologously expressed the gene successfully with a 3'-RACE combined with nest PCR strategy. Biological activity analyses indicated that the expressed peptide toxin, named latroeggtoxin-Ⅵ (LETX-Ⅵ), could inhibit Na⁺ channel currents in ND7/23 cells and promote dopamine release from PC12 cells, without obvious toxicity against Periplaneta americana and bacteria as well as fungi including Staphylococcus aureus and Candida albicans, demonstrating that LETX-Ⅵ is a mammal-specific neurotoxin with a potential application prospect in development of the tool reagents for neurobiological study and the drugs for treating related diseases.