title: Construction and identification of recombinant lentiviral vector containing HIV-1 Tat gene and its expression in 293T cells.
abstract: OBJECTIVE
  To construct a lentiviral vector expressing HIV-1 Tat and identify its expression in 293T cells.
  
  
  METHODS
  The gene fragment of HIV-1 Tat101 was subcloned to lentiviral transfer vector pHAGE-CMV-MCS-IZsGreen, which was named pHAGE-Tat. Then the constructed pHAGE-Tat was used to co-transfect the packing 293T cells, together with the packaging plasmids pMD2.G and psPAX2. The packaged viral particles designated LV-Tat were used to infect the 293T cells and the viral titer was calculated. The expression of HIV-1 Tat in 293T cells was confirmed using RT-PCR and western blot.
  
  
  RESULTS
  The recombinant lentiviral vector was successfully constructed and could express HIV-1 Tat in 293T cells. The virus titer was 5.73×10(6) ifu/ml.
  
  
  CONCLUSIONS
  The successfully constructed recombinant lentiviral vector makes a strong foundation for further exploring the possible role of HIV-1 Tat in the development of prostate cancer.
authors.affiliation: Department of Urology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
authors.forename: Bingbing
authors.initials: B
authors.lastname: Wei
authors.forename: Ninghan
authors.initials: N
authors.lastname: Feng
authors.forename: Feng
authors.initials: F
authors.lastname: Zhou
authors.forename: Chun
authors.initials: C
authors.lastname: Lu
authors.forename: Jiantang
authors.initials: J
authors.lastname: Su
authors.forename: Lixin
authors.initials: L
authors.lastname: Hua
keywords: HIV-1; Tat; prostate cancer
references.citation: Br J Cancer. 2000 Feb;82(3):718-25
references.pmid: 10682688
references.citation: J Virol. 2007 Mar;81(5):2401-17
references.pmid: 17151125
references.citation: Eur J Gynaecol Oncol. 2009;30(2):128-32
references.pmid: 19480239
references.citation: J Infect Dis. 1997 Jul;176(1):84-93
references.pmid: 9207353
references.citation: Adv Virus Res. 1999;52:139-232
references.pmid: 10384236
references.citation: Yale J Biol Med. 2006 Dec;79(3-4):115-22
references.pmid: 17940621
references.citation: Surv Ophthalmol. 2008 Jul-Aug;53(4):312-31
references.pmid: 18572051
references.citation: N Engl J Med. 1996 May 2;334(18):1168-72
references.pmid: 8602183
references.citation: Trends Microbiol. 2009 Mar;17(3):119-29
references.pmid: 19230674
references.citation: Postgrad Med J. 2008 Jul;84(993):372-7
references.pmid: 18716017
references.citation: J Natl Cancer Inst. 2008 Apr 16;100(8):528-9
references.pmid: 18398090
references.citation: Genome Res. 2009 Mar;19(3):438-51
references.pmid: 19208682
references.citation: J Virol. 1996 Nov;70(11):8151-4
references.pmid: 8892944
references.citation: Nat Med. 1995 Jul;1(7):707-8
references.pmid: 7585156
references.citation: Am J Dermatopathol. 2009 Aug;31(6):561-73
references.pmid: 19590418
references.citation: Curr Probl Dermatol. 2007;35:39-51
references.pmid: 17641489
references.citation: Annu Rev Pathol. 2009;4:399-415
references.pmid: 18928409
references.citation: Science. 1995 Apr 28;268(5210):582-3
references.pmid: 7725108
references.citation: J Infect Dis. 2004 Jan 1;189(1):15-20
references.pmid: 14702148
references.citation: N Engl J Med. 1998 Apr 2;338(14):948-54
references.pmid: 9521982
references.citation: Lab Invest. 2009 Jan;89(1):80-90
references.pmid: 19002111
references.citation: J Virol. 1996 Jan;70(1):549-58
references.pmid: 8523568
references.citation: Science. 1994 Dec 16;266(5192):1865-9
references.pmid: 7997879
references.citation: CA Cancer J Clin. 2007 Jan-Feb;57(1):43-66
references.pmid: 17237035
references.citation: Public Health Genomics. 2009;12(5-6):268-80
references.pmid: 19684440