title: [Construction and transduction of hNUDC shRNA interference lentiviral vector].
abstract: OBJECTIVE
  To construct a lentiviral vector carrying human nuclear distribution protein C (hNUDC)-specific shRNA (sh-NUDC-F) and knock down hNUDC expression in Dami cells by infection of the lentivirus.
  
  
  METHODS
  After labeling of green fluorescent protein (GFP),  sh-NUDC-F was cloned into lentiviral vector pRRL-cPPT-CMV-X-PRE-SIN,  and the high-quality plasmid was transfected into 293T cells to produce lentiviral particles by the calcium phosphate method. After high-speed centrifugation,  lentiviral particles were collected and determined for its titer. The high-titer lentiviral particles were then transduced into Dami cells. By detecting the expression of GFP in lentiviral vector using a microscope,  the transduction efficiency was readily figured out. And hNUDC protein level was detected by Western blotting.
  
  
  RESULTS
  hNUDC was totally knocked down by the efficient transduction of Dami cells with the lentivirus carrying  sh-NUDC-F.
  
  
  CONCLUSIONS
  Lentiviral vector  containing  sh-NUDC-F can be successfully packaged in 293T cells and then efficiently transduced into Dami cells to silence hNUDC gene.
authors: Department of Biology, Guangdong Medical College, Zhanjiang 524023, China. psf1975@sina.com
authors: Shi-feng
authors: SF
authors: Pang
authors: Xia
authors: X
authors: Liao
authors: Ke-qin
authors: KQ
authors: Zheng
authors: Ru-bin
authors: RB
authors: Zhou
authors: Peng
authors: P
authors: Liang
authors: Ding-guo
authors: DG
authors: Cao
authors: Guo-ping
authors: GP
authors: Zhang
authors: Wen-rong
authors: WR
authors: Li
mesh_terms: D018797:Cell Cycle Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D020868:Gene Silencing; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic
chemical_list: D018797:Cell Cycle Proteins; C512648:NUDC protein, human; D009687:Nuclear Proteins; D034741:RNA, Small Interfering