title: [Construct Mxi1 mutation gene expression vector from leukemia cell line KG1].
abstract: OBJECTIVE
  To construct the eukaryotic expression vector for Max interacting protein 1 (Mxi1).
  
  
  METHODS
  The full length cDNA of Mxi1 gene obtained from fetal lymphocyte and KG1 cells were inserted into plasmid pDs-red2-N1 respectively to generate pDs-red2-N1/Mri1 (wild/mutation type). Then the recombinant vector was transfected into Cos-7 cells via liposome. 48 hours post transfection, mRNA of Mri1 gene was detected by RT-PCR and Mxi1 protein expression was detected by flow cytometry and fluorescence microscope in the Cos-7 cells.
  
  
  RESULTS
  The eukaryotic expression vector of Mxi1 was constructed and transfected into eukaryotic cells successfully. The expression of red fluorescence protein in the transfected Cos-7 cells was observed under fluorescence microscope which implied the expression of Mxi1. The transfect efficiency of both wild and mutation type were in a high level in 3 days after transfected, which lasted to 6 d. RT-PCR amplified the total RNA extracted from the transfected Cos-7 cells could find increased mRNA level of Mxi1 gene.
  
  
  CONCLUSIONS
  We successfully constructed the eukaryote expression vector for Mri1 gene; Cos-7 cells transfected with the vector via liposome could express Mxi1 protein. These could be useful for the further study of the Myc gene modulation.
authors.affiliation: Department of Hematology, Key Laboratory of Heibei Institute of Hematology, 2nd Hospital of Hebei Medical University, Shijiazhuang 050000, China.
authors.forename: Xiao-Ling
authors.initials: XL
authors.lastname: Guo
authors.forename: Zhi-Yun
authors.initials: ZY
authors.lastname: Niu
authors.forename: Gai-Ling
authors.initials: GL
authors.lastname: Zhang
authors.forename: Feng-Lei
authors.initials: FL
authors.lastname: Yin
authors.forename: Ling
authors.initials: L
authors.lastname: Yang
authors.forename: Jin-Hai
authors.initials: JH
authors.lastname: Ren
authors.forename: Zuo-Ren
authors.initials: ZR
authors.lastname: Dong
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
authors.forename: Ling
authors.initials: L
authors.lastname: Pan
mesh_terms: D000818:Animals; D001483:Base Sequence; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics*; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D015973:Gene Expression Regulation, Leukemic; D005822:Genetic Vectors / Q000235:genetics*; D018257:Helix-Loop-Helix Motifs / Q000235:genetics; D006801:Humans; D007938:Leukemia / Q000473:pathology*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000235:genetics*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D025521:Tumor Suppressor Proteins; C079440:MXI1 protein, human