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	利用唐鱼雌激素受体构建环境雌激素重组酵母测评系统的研究; Recombinant yeast assay system for environmental estrogens mediated by estrogen receptor from Tanichthys albonubes
	陈美玲 ; 刘琳 ; 杨志兵 ; 武佳韵 ; 黄儒强 ; 李赞 ; 凌源智 ; 曾健辉 ; 马广智
	2014-05-07
关键词	唐鱼 环境雌激素 重组酵母评测系统 雌激素受体 β-半乳糖苷酶 Tanichthys albonubes environmental estrogens recombinant yeast assay estrogen receptor beta-galactosidase
英文摘要	利用唐鱼雌激素受体基因片段构建重组酵母,用以筛选环境雌激素类化学物质.实验先将唐鱼雌激素受体TErα基因片段插入载体PgAdT7中构建表达质粒P gAdT7/TErα,同时将雌激素效应元件(ErE)片段插入PMP206载体中构建报告质粒PMP206/ErE-lAC z;然后把表达质粒和报告质粒共转化到酵母AH109中,经筛选成功构建了由唐鱼雌激素受体调控的表达lAC z基因的重组酵母AHPTErα/ErE.结果表明,所构建的重组酵母AHPTErα/ErE在不同浓度17β-雌二醇(E2)的诱导下,β-半乳糖苷酶的活性呈现出明显的剂量-效应关系,其EC50为(0.521±0.700)nMOl·l-1.与dMSO对照组相比,重组酵母在17β-雌二醇诱导下有明显的β-半乳糖苷酶活性增强的现象.在不同浓度17α-乙炔基雌二醇(EE2)、壬基酚(nP)及其混合物、双酚A(bPA)、17β-雌二醇(E2)(阳性对照)的诱导下,重组酵母均呈现出剂量-效应关系,且灵敏度大小为E2>EE2>nP>bPA.结果表明,本研究成功构建了重组基因酵母测评系统,初步判定该重组酵母可应用于环境雌激素的筛选.; Yeast with recombinant estrogen receptor fragment of Tanichthys albonubes( terα) was used to screen the environmental estrogen-like compounds.terα and estrogen responsive element( ere) were cloned into pGADT7 and pMP206 respectively to obtain receptor( pGADT7 / terα) and reporter( p MP206 / ERE-Lac Z) constructs.The recombinant yeast AHpTERα / ERE was obtained by co-transformation of the receptor and reporter constructs,in which the expression of lac Z is regulated by the activity of terα.The assay sensitivity was evaluated by different concentrations of 17β-estradiol( E2) treatment.The results showed that the activity of beta-galactosidase induced by E2 was dose-dependent,and was significantly higher than that of DMSO control.The EC50 of E2 was( 0.521 ± 0.700) nmol·L- 1.The assay system is also sensitive to other estrogen compounds like 17α-ethynylestradiol( EE2),Bisphenol A( BPA) and nonylphenol( NP) technical mixtures dose-dependently,and the sensitivity of different chemicals ranked as E2> EE2> NP > BPA.The results showed that recombinant yeast assay system for environmental estrogens could be applied to screen the estrogens-like compounds in the environment.; 广东省科技计划项目(No.20120401)~~
语种	zh_CN
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/119326]
专题	生命科学－已发表论文
推荐引用方式 GB/T 7714	陈美玲,刘琳,杨志兵,等. 利用唐鱼雌激素受体构建环境雌激素重组酵母测评系统的研究, Recombinant yeast assay system for environmental estrogens mediated by estrogen receptor from Tanichthys albonubes[J],2014.
APA	陈美玲.,刘琳.,杨志兵.,武佳韵.,黄儒强.,...&马广智.(2014).利用唐鱼雌激素受体构建环境雌激素重组酵母测评系统的研究..
MLA	陈美玲,et al."利用唐鱼雌激素受体构建环境雌激素重组酵母测评系统的研究".(2014).

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