少棘蜈蚣钠通道毒素NTX-Ssm97的表达、纯化和鉴定  

Expression,Purification and Identification of Sodium Channel Toxins NTX-Ssm97 from Centipede Scolopendra Subspinipes

作　　者：尹世金[1] 李羽欣[2] 陆春兰[2] 邸智勇[3] 兰珍[2] 胡青兰[1] 

机构地区：[1]中南民族大学药学院,武汉430074 [2]中南民族大学生物医学工程学院,武汉430074 [3]武汉大学生命科学学院,武汉430071

出　　处：《中南民族大学学报（自然科学版）》2014年第2期49-53,共5页Journal of South-Central University for Nationalities：Natural Science Edition

基　　金：国家自然科学基金资助项目(81373379;30900239)

摘　　要：以同源序列比对法从湖北少棘蜈蚣毒腺转录组数据库中筛选到了一个新的毒素多肽基因NTX-Ssm97,其序列与Nav1.7抑制性多肽μ-SLPTX-Ssm6a高度同源.采用基因工程技术,构建了pGEX-4T-1-NTX-Ssm97原核表达载体,并利用E.coli Rosetta对毒素成熟肽的原核诱导表达和分离纯化.MALDI-TOF-MS质谱鉴定结果显示:纯化的NTX-Ssm97分子量与理论值基本吻合,表明少棘蜈蚣转录组数据库有助于筛选靶向Nav1.7钠通道的抑制性毒素多肽基因.A new toxin polypeptide gene of NTX-Ssm97 highly homologous to μ-SLPTX-Ssm6a, an inhibitory polypeptide of Nay 1.7 channels, was screened by homologous sequence alignment from the venomous transcriptome database of centipede Scolopendra subspinipes from Hubei Province. A prokaryotic expression vector pGEX-4T-1-NTX-Ssm97 was then constructed by genetic engineering techniques and the NTX-Ssm97 mature polypeptides were expressed and purified in E. coli Rosetta. MALDI-TOF-MS mass spectrometry showed that the molecular weights of purified NTX-Ssm97 were consistent with the theoretical value, suggesting that Seolopendra transcriptome database could contribute to the screening of the inhibitory peptide toxin genes for Navl. 7 channels.

关 键 词：少棘蜈蚣 转录组 NTX-Ssm97多肽 NAV1 7钠通道 基因工程 

分 类 号：TQ032.4[化学工程]