以绿色荧光蛋白为报告基因的环境砷离子生物检测体系的构建  

Constrution of method for detection of environmental arsenic based on green fluoresent protein as reporting gene

作　　者：胡春霞[1] 叶梦情[1] 岑益超[1] 易明花[1] 

机构地区：[1]绍兴文理学院元培学院生命科学系,浙江绍兴312000

出　　处：《环境与健康杂志》2013年第7期634-636,F0003,共4页Journal of Environment and Health

基　　金：绍兴文理学院重点科研项目

摘　　要：目的建立环境中砷的生物检测方法。方法根据GenBank中已发表的革兰阳性菌中参与砷抗性机制的操纵子序列R46邮，人工合成arsR全部序列（含小部分arsD基因序列）及其上游的启动子序列，以绿色荧光蛋白基因为报告基因，构建含砷离子特异性诱导启动子表达载体pET28a（＋）-ars-gfp，并采用感受态转化法将该重组表达载体转化人大肠埃希菌表达宿主菌EscherichiacoliBL21（DE3）中，构建砷离子检测菌株，并进行PCR和酶切鉴定。结果通过PCR和酶切鉴定，表明重组表达载体成功构建并转化人大肠杆菌EcoliBL21（DE3），绿色荧光蛋白报告基因在有砷离子（0．2～1．0μmol／L）存在的环境中得到表达。结论该菌株适用于环境中砷的生物检测。Objective A new method was developed to detect environmental arsenic, in which green fluorescent protein gene （GFP） and the sequences of arsenic-specific promoter and operon were ligated to form a new construct. Methods The ars region as well as the entire arsR gene （a small portion of arsD） and its upstream promoter were artificially synthesized based on the published sequence of the R46 ars operon, and were ligated into recombinant expression vector pET28a （＋） -gfp, gfp as reporting gene. The inducible expression vector named pET28a （＋） -ars-gfp were constructed successfully, and Escherichia coli BL21 （DE3） strain as the host. Results After PCR and double-enzyme cleavage, the recombinant strain was obtained and expressed as anticipated. Conclusion The new arsenic detection method established in this study by using bio-engineered construct is applicable to the determination of environmental arsenic.

关 键 词：砷 生物检测 绿色荧光蛋白 

分 类 号：R117[医药卫生—公共卫生与预防医学]