转基因表达有机磷水解酶(OPH)提高黄瓜降解蝇毒磷能力的初步研究  

Transgenic Expression of Organophosphorus Hydrolase(OPH)in Cucumber to Enhance the Detoxification of Coumaphos

作　　者：赵杰宏[1] 韩洁[2] 赵德刚[1] 

机构地区：[1]贵州大学农业生物工程省重点实验室教育部绿色农药与农业生物工程重点实验室,贵州贵阳550025 [2]贵阳中医学院基础部微生物教研室,贵州贵阳550002

出　　处：《江苏农业学报》2010年第1期182-186,共5页Jiangsu Journal of Agricultural Sciences

基　　金：国家科技支撑计划(2007BAD59B00);国际科技合作项目(2007DFA31260)

摘　　要：为快速提高黄瓜降解有机磷农药残留的能力，选取能广泛降解有机磷农药的有机磷水解酶（OPH）为表达蛋白，构建CaMV35S启动子驱动有机磷水解酶基因（opd）的植物表达载体pSOP，经双酶切证明重组载体含有插入位置和方向皆正确的目的片段。通过农杆菌介导遗传转化黄瓜子叶节，诱导和筛选出9株抗性苗，获得3株GUS染色和PCR阳性转化苗。酶活性分析表明，转基因黄瓜降解蝇毒磷能力是对照（非转基因株）的4．7～9．7倍，最高达到7．8μmol／（mg·min），有助于加快降解黄瓜中的有机磷农药残留，为食品安全研究提供借鉴。In order to enhance the degrading effect of cucumber on organophosphorus pesticides residues, a plant expression vector of opd driven by CaMV 35S promoter was constructed. The detection results of restriction enzymes digestions indicated that the vector held the objective gene sequence of opd and the position or direction inserted were correct. The opd gene was transformed into cucumber explants. GUS staining and PCR confirmed that the opd gene had been integrated into the cucumber genome. The degradation of transgenic cucumbers were 4. 7-9. 7 times as much as that of the non- transformed one, with the maximum degradation up to 7.8 μmoll （ mg·min）.

关 键 词：生物修复 遗传转化 有机磷水解酶(OPH) 农药残留 

分 类 号：Q78[生物学—分子生物学] X592[环境科学与工程—环境工程]