摘要
根据已克隆的鲢鱼(Hypophthalmichthysmolitrix)微囊藻毒素去毒酶cDNA全序列设计特异引物,利用PCR方法获得鲢鱼微囊藻毒素去毒酶基因编码区,将该编码区与绿色荧光蛋白连接,分别构建融合表达载体pEGFP-N1-sGST和双顺反子表达载体pIRES2-EGFP-sGST。利用脂质体转染法将融合表达载体pEGFP-N1-sGST转染Hela细胞,60h后检测到绿色荧光基因表达;通过显微注射,将双顺反子表达载体pIRES2-EGFP-sGST注入斑马鱼(Daniorerio)受精卵,获得了转鲢鱼微囊藻毒素去毒酶基因斑马鱼,从而构建了微囊藻毒素去毒酶转基因模型。上述2种转基因模型的成功构建为进一步研究鲢鱼、鳙鱼(Aristichthysnobilis)、罗非鱼(Oreochromisnilotica)等淡水鱼类微囊藻毒素去毒酶基因调控元件、去毒分子机理及研发转基因鲢鱼、鳙鱼、罗非鱼等微囊藻毒素高效生物去毒器奠定了基础。
cDNA fragment encoding complete sequence of silver carp （Hypophthalmichthys molitrix） microcystin-detoxifizyme was subcloned to construct a lhsing gene expression vector pEGFP-NI-sGST, and a dicistronic expression vector plRES2-EGFP-sGST. Using lipofectin method, the fusing gene expression vector pEGFP-NI-sGST was transfected into Hela cells. The emission of fluorescence was observed after 60 hours. Through microinjection, the dicistronic expression vector plRES2-EGFP-sGST was delivered into embryos of zebrafish （Danio rerio）, and the silver carp microcystin-detoxifizyme transgenic zebrafish was obtained. The success of the two transgenic models makes it possible to further analyze the cis-regulatory elements of the microcystin-detoxifizyme gene, and to develop a strain of transgenic planktivorous fish, used as a high-efficient biodetoxicator for microcystin in lakes and reservoirs.
出处
《生态科学》
CSCD
2006年第1期25-27,31,共4页
Ecological Science
基金
广东省水文局蓝藻重点项目
广东省自然科学基金项目(031886)
教育部留学回国人员科研启动基金项目
