title: [Cloning, eukaryotic expression and celluar localization of cysteine and glycine-rich protein 1].
abstract: OBJECTIVE
  To construct an eukaryotic expression vector of pCMV-FLAG-CRP1 and investigate the cellular localization of cysteine and glycine-rich protein 1 (CRP1) in human embryonic kidney 293T, hepatocellular carcinoma HepG2 and breast cancer ZR75-1 cells.
  
  
  METHODS
  Human CRP1 gene was obtained from mammary cDNA library by PCR and cloned into the pCMV-Tag2B vector. Human 293T, HepG2 and ZR75-1 cells were transfected with the recombinant plasmid pCMV-FLAG-CRP1; the expression was detected by Western blotting and the cellular localization was observed under a fluorescence microscope.
  
  
  RESULTS
  CRP1 eukaryotic expression vector labeled with FLAG tag was successfully constructed as demonstrated by double enzyme digestion and gene sequencing. Western blotting showed that CRP1 protein was expressed in FLAG-CRP1 transfected 293T, HepG2 and ZR75-1 cells, and fluorescence microscopy revealed that CRP1 protein was localized in both cytoplasm and nucleus of the three cell lines, and the former signal was stronger than the latter.
  
  
  CONCLUSIONS
  The eukaryotic expression vector of pCMV-FLAG-CRP1 is successfully constructed, and the cellular location of CRP1 is further identified in the 293T, HepG2 and ZR75-1 cell lines, which will contribute to the further study on the function of CRP1.
authors.affiliation: Beijing Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Jiahong
authors.initials: J
authors.lastname: Liu
authors.forename: Xiaojie
authors.initials: X
authors.lastname: Xu
authors.forename: Jing
authors.initials: J
authors.lastname: Fu
authors.forename: Zhongyi
authors.initials: Z
authors.lastname: Fan
authors.forename: Chaohui
authors.initials: C
authors.lastname: Lv
authors.forename: Juming
authors.initials: J
authors.lastname: Lu
authors.forename: Jianhua
authors.initials: J
authors.lastname: Zhu
authors.forename: Qinong
authors.initials: Q
authors.lastname: Ye
mesh_terms: D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D003001:Cloning, Molecular; D015870:Gene Expression; D005822:Genetic Vectors; D057809:HEK293 Cells; D056945:Hep G2 Cells; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D060588:LIM Domain Proteins / Q000235:genetics* / Q000378:metabolism*; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: C486181:CRIP1 protein, human; D002352:Carrier Proteins; D060588:LIM Domain Proteins; D011993:Recombinant Fusion Proteins