Gene cloning, heterologous expression and activity identification of latroeggtoxin-

• Published in: Sheng wu gong cheng xue bao = Chinese journal of biotechnology Vol. 37; no. 2; p. 635
• Main Authors: Yan, Shuai, Tang, Xiaochao, Yu, Dianmei, Wang, Haiyan, Meng, Wenwen, Tang, Pingping, Wang, Xianchun
• Format: Journal Article
• Language: Chinese
• Published: China 25-02-2021
• Subjects: Animals; Arthropod Proteins - genetics; Black Widow Spider - genetics; Cloning, Molecular; Rats; Spider Venoms - genetics; Transcriptome; nest PCR; peptide; biological activity; black widow spider; latroeggtoxin; rapid-amplification of CDNA ends
• ISSN: 1872-2075

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 aureu