Gene cloning, heterologous expression and activity identification of latroeggtoxin-Ⅵ.

Shuai Yan; Xiaochao Tang; Dianmei YU; Haiyan Wang; Wenwen Meng; Pingping Tang; Xianchun Wang

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Gene cloning, heterologous expression and activity identification of latroeggtoxin-Ⅵ.

Author: Shuai YAN ¹ ; Xiaochao TANG ¹ ; Dianmei YU ¹ ; Haiyan WANG ¹ ; Wenwen MENG ¹ ; Pingping TANG ¹ ; Xianchun WANG ¹
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1. Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, Hunan, China.
Publication Type:Journal Article
Keywords: biological activity; black widow spider; latroeggtoxin; nest PCR; peptide; rapid-amplification of CDNA ends
MeSH: Animals; Arthropod Proteins/genetics*; Black Widow Spider/genetics*; Cloning, Molecular; Rats; Spider Venoms/genetics*; Transcriptome
From: Chinese Journal of Biotechnology 2021;37(2):635-645
CountryChina
Language:Chinese
Abstract: 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.