title: [Construction of Trim6 eukaryotic expression vector and its expression in HEK293 cells].
abstract: OBJECTIVE
  To construct the recombinant eukaryotic expression vector pcDNA3.1 (+)-Trim6, and observe its expression in HEK293T cells in vitro.
  
  
  METHODS
  The total RNA was isolated from HeLa cells. After amplification with reverse transcription polymerase chain reaction (RT-PCR), the target sequences were cloned into the pcDNA3.1(+). The recombinant vector was confirmed by restriction enzyme digestion, PCR and sequencing. Then it was transfected into HEK293T cells.After 24 hours, the Trim6 expression was detected by Western blot.
  
  
  RESULTS
  The results of the restriction enzyme digestion, PCR and sequencing confirmed the vector was constructed successfully, and it can express Trim6 protein in HEK293T cells.
  
  
  CONCLUSIONS
  The vector is constructed successfully, which establishes the foundation for future research on the effect of Trim6.
authors: Experiment Center of Clinical Medicine, Affiliated Hospital of Binzhou Medical University, Binzhou 256603, China.
authors: Da-Kang
authors: DK
authors: Sun
authors: Xin-Ye
authors: XY
authors: An
authors: Feng-Ai
authors: FA
authors: Hu
authors: Cai-Yu
authors: CY
authors: Li
authors: Jing
authors: J
authors: Zheng
mesh_terms: D001483:Base Sequence; D018076:DNA, Complementary; D005822:Genetic Vectors / Q000235:genetics*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D000070584:Tripartite Motif Proteins; D044767:Ubiquitin-Protein Ligases / Q000235:genetics* / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D009124:Muscle Proteins; D000070584:Tripartite Motif Proteins; C440908:TRIM63 protein, human; D044767:Ubiquitin-Protein Ligases