蝎昆虫神经毒素及其基因研究进展

蝎昆虫神经毒素是一类对昆虫有专一性麻痹致死作用，而对哺乳动物无害或毒性很小的蛋白。因此分离和表达蝎昆虫神经毒素基因，有利于开发研究新型高效生物杀虫剂，并为把该基因导入植物培育新抗虫品种提供必要的基础。另外，此类蝎昆虫神经毒素还有利于研究电压依从性离子通道结构与功能的相互关系。对蝎昆虫神经毒素的研究已成为国内外生物科学工作者关注的课题之一。本文综述了近几年来蝎昆虫神经毒素及其基因的研究进展概况。     

Cloning and sequencing of two depressant insect selective neurotoxin cDNAs from Buthus martensii Karsch

During evolution, scorpions have developed the ability to produce a series of toxins. Scorpion toxins, which are reserved in terminal segments of scorpion, serve as the arms for predation and self-defence. They have also been used as medicine for some human sickness. As far as the targets that they act on are concerned, these toxins can be divided     

A型肉毒神经毒素纯化及其免疫原性研究

采用阴离子交换柱层析法和凝胶过滤柱层析法纯化A型肉毒神经毒素复合物,通过SDS-PAGE法分析不同纯化条件下目标物的纯度和完整性,并以纯化物免疫BALB/c小鼠研究其免疫原性.结果显示,离子交换柱层析法纯化效果优于凝胶过滤柱层析法.     

Polyclonal antibody against an insect excitatory toxin BmKIT from Buthus Martensii karsch and detection of BmKIT expressed in transgenic cotton

An insect excitatory toxin gene from Buthus martensii Karseh （BmKIT） was cloned into the expression vector, pET-28a. BmKIT was expressed as inclusion bodies in Eseherichia coli BL21 （DE3） host cells. The authenticity of in vitro expressed protein was confirmed by Western blot. The inclusion body protein band in SDS-PAGE was excised and the protein, BmKIT, was extracted. Polyelonal antibodies to the purified protein were raised in rabbits. The antibody reacted specifically with the expressed BmKIT and was used to quantify its presence in transgenic cotton.     

虎纹捕鸟蛛（Selenocosimia huwena）粗毒对蟾蜍神经肌肉接头传递的影响

１０－５～１０－４ｇ／ｍｌ虎纹捕鸟蛛粗毒对蟾蜍坐骨神经肌肉标本有阻遏作用，１×１０－４ｇ／ｍｌ粗毒发生阻断的时间为３８ｍｉｎ，但不改变神经干动作电位，不影响肌肉对直接刺激的反应。阻断后肌肉对间接强直刺激仍可发生强直收缩；粗毒对蟾蜍坐骨神经肌肉标本的阻断作用表现为：初期４～５０秒的收缩增强继而转向抑制；粗毒可对抗箭毒的阻断作用；蟾蜍在体坐骨神经肌肉标本可被粗毒阻断，但阻断后可以恢复；粗毒不改变蛙腹直肌对Ａｃｈ的敏感性，粗毒中含有拟胆碱成份。箭毒可以阻止粗毒对蛙腹直肌的拟胆碱效应；该粗毒对蟾蜍坐骨神经肌肉标本的阻遏作用发生在神经肌肉接头，粗毒中含有拟胆碱组份，此组份可能通过后膜去极化阻断神经肌肉接头。     

Three-dimensional solution structure of w-conotoxin SO3 determined by ~1H NMR

Cone snails (Conus) elaborate a series of conotoxin (CTX) peptides in their venoms to paralyze their prey. Many of these toxins specifically block ion channels in vertebrate and invertebrate neuromuscular systems[1]. The pattern of Cys residues is quite conserved in most CTXs and can be arranged in three major frameworks: CCCC (a-CTX), CCCCCC (m-CTX), CCCCCC (w-CTX). These peptides have been used as important tools in receptor and ion channel studies[2]. Among these toxins, w-CT…     

Baculovirus-mediated expression of a Chinese scorpion neurotoxin improves insecticidal efficacy

An Buthus martensii Karsch Insect Toxin （BmK IT ） gene was inserted into the genome of Autographa californica multicapsid nucleopolyhedrovirus （AcMNPV） to construct a recombinant baculovirus, AcMNPV-BmK IT. The expression of BmK IT was confirmed using RT-PCR, dot blot and SDS-PAGE analysis. Dose-lethal time responses to Spodoptera exigua larvae were compared between wild-type baculovirus AcMNPV and recombinant virus AcMNPV-BmK IT. At the concentration of 1 × 10^7 PIBs/mL, the median lethal time of recombinant baculovirus （LT50 = 73.6 h） on third instar S. exigua larvae showed an improvement of 13.2% over the efficacy of wild type virus （LT50 = 84.8 h） during a 192 h infection.     

A型肉毒杆菌毒素的基因测序与分析

对国内A型肉毒杆菌毒素基因测序,分析了解毒素基因结构并推测其氨基酸序列。提取肉毒杆菌总DNA,利用自行设计的引物对目的基因进行PCR扩增,把目的基因导入E.Coli JM109,筛选含目的基因的阳性克隆菌进行测序和分析,从而了解毒素基因结构并推测其氮基酸序列。对A型肉毒杆菌毒素基因的核苷酸序列成功测序并与GenBank中的相应序列比较,其序列同源性为96％。推测其氨基酸序列同源性为100％。成功地对A型肉毒杆菌毒素的基因进行了测序,这为肉毒杆菌毒素中毒的快速诊断,以及进一步探求以基因工程方法生产此毒素奠定基础。