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逯晓波, 于涛, 刘艳华, 肖莎, 靳翠红. ERCC1密码子118单核苷酸多态性转染细胞模型建立[J]. 中国公共卫生, 2011, 27(10): 1247-1249. DOI: 10.11847/zgggws-2011-27-10-14
引用本文: 逯晓波, 于涛, 刘艳华, 肖莎, 靳翠红. ERCC1密码子118单核苷酸多态性转染细胞模型建立[J]. 中国公共卫生, 2011, 27(10): 1247-1249. DOI: 10.11847/zgggws-2011-27-10-14
LU Xiao-bo, YU Tao, LIU Yan hua, . Establishment of cell transfected model for ERCC1 codon 118 single nucleotide polymorphisms[J]. Chinese Journal of Public Health, 2011, 27(10): 1247-1249. DOI: 10.11847/zgggws-2011-27-10-14
Citation: LU Xiao-bo, YU Tao, LIU Yan hua, . Establishment of cell transfected model for ERCC1 codon 118 single nucleotide polymorphisms[J]. Chinese Journal of Public Health, 2011, 27(10): 1247-1249. DOI: 10.11847/zgggws-2011-27-10-14

ERCC1密码子118单核苷酸多态性转染细胞模型建立

Establishment of cell transfected model for ERCC1 codon 118 single nucleotide polymorphisms

  • 摘要: 目的 建立切除修复交叉互补集团1(ERCC1)基因单核苷酸多态性(SNP)转染细胞模型,为体外研究基因多态性功能提供平台。方法 Gateway定向克隆技术构建ERCC1表达载体,包含不同ERCC1 codon 118 SNP表达载体转染中国仓鼠卵巢细胞(CHO)缺陷型UV20细胞,蛋白免疫印迹法(western blot)检测ERCC1蛋白表达,焦磷酸短序列测序PSQ检测ERCC1 118 SNP基因型。结果 酶切鉴定及测序分析表明ERCC1表达载体构建正确,各转染细胞表达绿色荧光蛋白并可在G418培养基选择生长;ERCC1在UV20细胞中表达分子量为38 kDa蛋白,ERCC1 SNP基因型正确,2种转染细胞ERCC1的SNP基因型不同,分别为C/C或T/T。结论 ERCC1 codon 118 SNP细胞转染模型构建成功,为体外研究ERCC1 codon 118 SNP不同基因型提供了实验技术平台。

     

    Abstract: Objective To establish a transfected cell model of excision repair cross complementation group 1(ERCC1) codon 118 single nucleotide polymorphisms(SNP)and to provide a valid platform for ERCC1 codon 118 SNP function study.Methods ERCC1 expressed vector was constructed by GatewayTM clone technique.The expression vectors including different ERCC1 codon 118 SNP genotypes were transfected to Chinese hamster ovary(CHO)cell line defective mutant UV20.ERCC1 protein expression was detected by western blot.ERCC1 codon 118 SNP genotype was tested by PSQTM.Results ERCC1 expression vectors were established through GatewayTM clone technique and confirmed by restrict enzyme and genotype analysis.All transfected cell line can grow in G418 DMEM medium and express green fluorecent protein,ERCC1 protein and genotypes were also confirmed by western bolt and PSQTM.Conclusion The transfected cell model related to ERCC1 codon 118 SNPs was established and could be adopted in the research of single nucleotide polymorphisms of DNA damage repair gene in vitro.

     

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