title: Feeding recombinant E. coli with GST-mBmKTX fusion protein increases the fecundity and lifespan of Caenorhabditis elegans.
abstract: Scorpion venom could be a useful treatment for a variety of diseases, such as cancer, epilepsy and analgesia. BmKTX is a polypeptide extracts from scorpion venom (PESV), which have attracted much attention from researchers in recent years. mBmKTX is a mutant polypeptide according to the amino acid sequence of BmKTX. We expressed it with the vector pGEX-4T-1 in Escherichia coli, and Caenorhabditis elegans were used as the animal model and fed with the strains. In this study, the expression of pGEX-mBmKTX was analyzed by SDS-PAGE, and GST-mBmKTX purified from pGEX-mBmKTX as a glutathione S-transferase (GST)-tagged fusion protein is approximately 30kDa. The secondary structure prediction shows that mBmKTX is mainly composed of approximately 13% β-sheet and 86% loop. A food clearance assay and brood size assay indicated that the worms fed pGEX-mBmKTX ate more and had greater fecundity than those fed the empty vector. A lifespan analysis demonstrated that mBmKTX could significantly prolong the lifespan of C. elegans, with an increase of 22.5% compared with the control. Behavioral assays confirmed that mBmKTX had no influence on the locomotion of C. elegans. In addition, microarray analysis and quantitative real-time PCR demonstrated that there are 320 differentially expressed genes, 182 of which are related to reproduction, growth and lifespan. In conclusion, the data suggested that mBmKTX has potential utility for increasing fecundity and animal survival.
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, and School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Xu
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, and School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Yajie
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, and School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Wan
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, and School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, and School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China; Guangdong Province Key Laboratory for Biotechnology Drug Candidates, School of Biosciences and Biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou 510006, China.
authors.forename: Zebo
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, and School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Yongmei
authors.initials: Y
authors.lastname: Liu
authors.affiliation: School of Life Science, Wuhan University, Wuhan 430071, China.
authors.forename: Yingliang
authors.initials: Y
authors.lastname: Wu
authors.affiliation: School of Life Science, Wuhan University, Wuhan 430071, China.
authors.forename: Zongyun
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, and School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China. Electronic address: lx@whu.edu.cn.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D017173:Caenorhabditis elegans / Q000187:drug effects / Q000235:genetics; D029742:Caenorhabditis elegans Proteins / Q000096:biosynthesis; D004926:Escherichia coli / Q000235:genetics; D005298:Fertility / Q000187:drug effects / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D005982:Glutathione Transferase / Q000235:genetics* / Q000494:pharmacology; D008136:Longevity / Q000187:drug effects / Q000235:genetics; D046228:Microarray Analysis; D050505:Mutant Proteins / Q000235:genetics* / Q000494:pharmacology; D010455:Peptides / Q000235:genetics* / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology; D012604:Scorpion Venoms / Q000235:genetics* / Q000494:pharmacology
chemical_list: D029742:Caenorhabditis elegans Proteins; C109066:KTX toxin, Buthus; D050505:Mutant Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; D012604:Scorpion Venoms; D005982:Glutathione Transferase
keywords: C. elegans; Fecundity; Lifespan; Microarray; mBmKTX