Feeding recombinant E. coli with GST-mBmKTX fusion protein increases the fecundity and lifespan of Caenorhabditis elegans.
Feeding recombinant E. coli with GST-mBmKTX fusion protein increases the fecundity and lifespan of Caenorhabditis elegans.
复制标题
用 GST-mBmKTX 融合蛋白喂养重组大肠杆菌可增加秀丽隐杆线虫的繁殖力和寿命
DOI：
10.1016/j.peptides.2017.01.003
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发表时间：
2017
期刊：
影响因子：
3
通讯作者：
XinLiu
中科院分区：
文献类型：
--
作者：
Jie Xu;Yajie Jiang;Lu Wan;Qi Wang;Zebo Huang;Yongmei Liu;Yingliang Wu;Zongyun Chen;XinLiu
蝎毒可能是一种有用的治疗多种疾病，如癌症，癫痫和镇痛。BmKTX是一种蝎毒多肽提取物，近年来引起了研究者的广泛关注。mBmKTX是根据BmKTX的氨基酸序列的突变多肽。将其与载体pGEX-4 T-1在大肠杆菌中进行表达，并以秀丽隐杆线虫为动物模型，进行了实验研究。在本研究中，通过SDS-PAGE分析pGEX-mBmKTX的表达，并且从pGEX-mBmKTX纯化的GST-mBmKTX作为谷胱甘肽S-转移酶（GST）标记的融合蛋白约为30 kDa。二级结构预测表明，mBmKTX主要由约13%的β-折叠和86%的环状结构组成。食物清除试验和产卵量试验表明，喂食pGEX-mBmKTX的蠕虫比喂食空载体的蠕虫吃得更多，具有更高的繁殖力。寿命分析表明mBmKTX能显著延长C.秀丽隐杆线虫，比对照组增加22.5%。行为学实验证实mBmKTX对C.优雅的此外，基因芯片分析和实时荧光定量PCR显示有320个差异表达基因，其中182个与生殖、生长和寿命相关。总之，数据表明mBmKTX具有增加繁殖力和动物存活的潜在效用。
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 30 kDa. 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.