GFP遗传改造四膜虫细胞的构建及其对重金属污染的荧光响应

发布时间：2018-06-08 13:12

本文选题：金属硫蛋白 + 绿色荧光蛋白　；参考：《太原理工大学学报》2017年06期

【摘要】：为了获得可用于检测环境中多种重金属污染的遗传改造生物细胞,将具有重金属诱导特性的金属硫蛋白基因MTT1启动子和外源报告基因绿色荧光蛋白gfp的融合序列,通过基因枪法转染嗜热四膜虫细胞。经同源重组和巴龙霉素抗性筛选,获得稳定的GFP遗传改造四膜虫细胞株。该细胞株经Cd~(2+)诱导,MTT1启动子可启动外源基因gfp正确表达,在蓝光激发下可发绿色荧光。GFP遗传改造四膜虫细胞对重金属离子的荧光响应效应由大到小依次为Cd~(2+),Cr~(6+),Hg~(2+);响应浓度范围和具有剂量-效应关系的浓度范围由大到小依次为Cd~(2+),Hg~(2+),Cr~(6+);最低检测限由大到小依次为Cr~(6+),Hg~(2+),Cd~(2+),而对Cu~(2+)无荧光响应效应。因此,GFP遗传改造四膜虫可作为环境中Cd~(2+)、Hg~(2+)和Cr~(6+)污染的生物检测细胞。
[Abstract]:In order to obtain genetically modified biological cells which can be used to detect the contamination of many heavy metals in the environment, the fusion sequences of MTT1 promoter of metallothionein gene and green fluorescent protein gfp of exogenous reporter gene were obtained. Tetrahymena thermophilus cells were transfected by gene gun method. After homologous recombination and baromycin resistance screening, stable GFP genetically modified Tetrahymena cell lines were obtained. This cell line was induced by CDT-1) to initiate the correct expression of exogenous gene gfp. The fluorescence response of Tetrahymena cells to heavy metal ions can be transformed by green fluorescence. GFP under blue light. The order of fluorescence response to heavy metal ions is from large to small, the response concentration range and the concentration range with dose-effect relationship are from large to small. The lowest detection limit from large to small is CrGN 2 / HgCU, and the lowest detection limit is CrP6 / HgCU 2 / 2, and no fluorescence response is found to Cuzhuan2) in order of the lowest detection limit of CD / C ~ (2 +) and Cr ~ (2 +) / C ~ (2 +), and the lowest detection limit is Cr ~ (6) HG ~ (2 +) ~ (2). Therefore, the GFP genetic transformation of Tetrahymena can be used as a bioassay cell contaminated by CD ~ (2 +) H _ (G) ~ (2) and Cr ~ (3 +) ~ (6) in the environment.
【作者单位】：太原理工大学信息工程学院;太原理工大学化学化工学院;
【基金】：国家自然科学青年基金资助项目(31501124) 山西省基础研究青年基金资助项目(201502119)
【分类号】：X174

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