title: [Construction and identification of a recombinant lentivirus harboring small interfering RNA targeting mouse CD99 antigen-like 2 gene].
abstract: OBJECTIVE
  To construct a recombinant lentivirus harboring RNA interference sequence targeting mouse CD99 antigen-like 2 (mCD99L2) gene and observe its infection efficiency of 293FT cells.
  
  
  METHODS
  Four pairs of small interfering RNAs (siRNAs) targeting mCD99L2 cDNA were designed, synthesized and linked to the lentivirus vector SD1259 to construct the lentivirus shuttle plasmids. After sequencing, the 4 lentivirus shuttle plasmids were transfected into 293FT cells in the presence of packaging plasmids. Forty-eight hours later, the supernatant was collected and the titer and infection efficiency of the recombinant lentivirus were determined according to the expression of the reporter gene enhanced green fluorescent protein (EGFP) under fluorescent microscope.
  
  
  RESULTS
  DNA sequencing demonstrated that mCD99L2 siRNAs were successfully cloned to the lentiviral vector SD1259. The titer of concentrated virus was 1x10(7)/ml in the supernatant of the infected cells.
  
  
  CONCLUSIONS
  The recombinant lentivirus containing siRNA targeting mCD99L2 gene has been successfully constructed, which provide the basis for future establishment of visualized cell model and animal model of Hodgkin's lymphoma.
authors.affiliation: Department of Pathology, Nanfang Hospital, Southern Medical University, and Key Lab for Molecular Tumor Pathology of Guangdong Province, Southern Medical University, Guangzhou 510515, China. zhanggong7446@163.com
authors.forename: Gong
authors.initials: G
authors.lastname: Zhang
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
authors.forename: Xiao-yan
authors.initials: XY
authors.lastname: Chen
authors.forename: Wei-yi
authors.initials: WY
authors.lastname: Fang
authors.forename: Hui-ling
authors.initials: HL
authors.lastname: Li
authors.forename: Zuo-ping
authors.initials: ZP
authors.lastname: Huang
authors.forename: Hong-juan
authors.initials: HJ
authors.lastname: Chu
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Tong
authors.initials: T
authors.lastname: Zhao
mesh_terms: D000071443:12E7 Antigen; D000818:Animals; D015703:Antigens, CD / Q000235:genetics*; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006689:Hodgkin Disease / Q000473:pathology*; D016086:Lentivirus / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics; D014407:Tumor Cells, Cultured
chemical_list: D000071443:12E7 Antigen; D015703:Antigens, CD; C522605:Cd99 protein, mouse; D018076:DNA, Complementary; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins