title: [Construction of mic2/CD99 gene vector and its transfection in Hodgkin lymphoma L428 cell line].
abstract: OBJECTIVE
  To construct a eukaryotic expression vector of CD99 gene for transfection into Hodgkin lymphoma L428 cells.
  
  
  METHODS
  The full-length cDNA of CD99 gene was amplified from Jurkat cells by RT-PCR and cloned into the pcDNA3.1(+) vector and transfected into L428 cell line using Lipofextamine 2000. The sequence of CD99 mRNA in the transfected cells was confirmed by restriction endonuclease digestion and DNA sequencing, and the expression of CD99 protein was identified using immunocytochemistry.
  
  
  RESULTS
  A gene fragment of 558 bp was amplified from the transfected cells and the sequence was verified by DNA sequencing. Immunocytochemistry identified the presence of CD99 expression in the transfected cells.
  
  
  CONCLUSIONS
  A eukaryotic expression vector pcDNA3.1(+)-CD99 is successfully constructed and stably expressed in L428 cell line.
authors.affiliation: Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. zuoping68@126.com
authors.forename: Zuo-ping
authors.initials: ZP
authors.lastname: Huang
authors.forename: Ying
authors.initials: Y
authors.lastname: He
authors.forename: Xin-hua
authors.initials: XH
authors.lastname: Zhou
authors.forename: Xi-qing
authors.initials: XQ
authors.lastname: Han
authors.forename: Zi-Qun
authors.initials: ZQ
authors.lastname: Wu
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Zong-Hua
authors.initials: ZH
authors.lastname: Wen
authors.forename: Tong
authors.initials: T
authors.lastname: Zhao
mesh_terms: D000071443:12E7 Antigen; D015703:Antigens, CD / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D015815:Cell Adhesion Molecules / Q000096:biosynthesis* / Q000235:genetics; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D005822:Genetic Vectors / Q000235:genetics*; D006689:Hodgkin Disease / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D019169:Jurkat Cells; D008969:Molecular Sequence Data; D014162:Transfection
chemical_list: D000071443:12E7 Antigen; D015703:Antigens, CD; C061828:CD99 protein, human; D015815:Cell Adhesion Molecules