title: [Construction of eukaryotic expression vector of human CD34 and establishment of stably transfected cell line].
abstract: OBJECTIVE
  To construct eukaryotic expression vector of human CD34 and transfect it to 3T3 cells so as to establish stably transfected 3T3 cell line.
  
  
  METHODS
  The RNA was extracted from KG1a. CD34 gene was amplified by RT-PCR. With the double-enzyme digestion, CD34 gene was cloned into pCI-neo eukaryotic expression vector, yielding pCI-CD34. The pCI-CD34 was transfected into 3T3 cell by electroporator. Stably transfected 3T3 cell line was established, and the CD34 expression in the transfected cells was detected by RT-PCR and FACS.
  
  
  RESULTS
  The eukaryotic expression vector pCI-CD34 was constructed, and stably transfected 3T3 cell line was established.
  
  
  CONCLUSIONS
  Construction of eukaryotic expression vector of CD34 and the establishment of stably transfected 3T3 cell line are helpful to preparation of anti-CD34 mAbs and further functional study of CD34.
authors.affiliation: Union Stem Cell & Gene Engineering Co., Ltd., Tianjin 300384, China. shilin0609@126.com
authors.forename: Lin
authors.initials: L
authors.lastname: Shi
authors.forename: Hao
authors.initials: H
authors.lastname: Qu
authors.forename: Hong-qin
authors.initials: HQ
authors.lastname: Liu
authors.forename: Hai-xia
authors.initials: HX
authors.lastname: Gao
authors.forename: Wei-bin
authors.initials: WB
authors.lastname: Zhu
mesh_terms: D016475:3T3 Cells; D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D005822:Genetic Vectors / Q000138:chemical synthesis* / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014162:Transfection
chemical_list: D018952:Antigens, CD34