title: [Construction of a lentiviral vector carrying TRAIL gene and its infection efficiency to lymphoma cells in vitro].
abstract: This study was purposed to construct a lentiviral vector carrying the TNF-related apoptosis-inducing ligand (TRAIL) gene and investigate its infection efficiency to several lymphoma cells lines. A pGM-T-TRAIL vector was constructed by inserting the cDNA segment derived from TRAIL mRNA into the cloning vector pGM-T, which was then inserted into the lentiviral vector pWPI. The recombinant lentiviral vector plenti-TRAIL was produced by transfecting 293T cells with pWPI-TRAIL, packaging plasmid Δ8.2, and envelope plasmid pCMV-VSVG and then harvested from the culture supernatant. Infection efficiency was measured in several lymphoma cell lines by live cell GFP fluorescence, while TRAIL expression was assessed by RT-PCR and Western blot. The results showed that the enzyme cut identification and sequencing demonstrated the successful construction of both pGM-T-TRAIL and pWPI-TRAIL. The results of testing drop showed that the concentration of the restructured lentiviral plenti-TRAIL reached 10(9) IU/ml. Comparison of infection efficiency revealed that YTS cells were more likely to be infected than DOHH2 or Jurkat cells (P < 0.05). Finally, RT-PCR and Western blot showed that lymphoma cells infected with plenti-TRAIL were able to efficiently express the TRAIL mRNA and protein. It is concluded that the lentiviral vector pWPI-TRAIL is successfully constructed and the recombinant lentiviral plenti-TRAIL is manufactured. The plenti-TRAIL vector is able to infect several lymphoma cell lines, and the infected lymphoma cells can effectively express TRAIL genes.
authors.affiliation: Department of Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
authors.forename: Xiao-Rui
authors.initials: XR
authors.lastname: Fu
authors.forename: Xu-Dong
authors.initials: XD
authors.lastname: Zhang
authors.forename: Chen
authors.initials: C
authors.lastname: Zhang
authors.forename: Lu
authors.initials: L
authors.lastname: Zhao
authors.forename: Bian-Hong
authors.initials: BH
authors.lastname: Wang
authors.forename: Ming-Zhi
authors.initials: MZ
authors.lastname: Zhang
mesh_terms: D001483:Base Sequence; D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008223:Lymphoma / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D053221:TNF-Related Apoptosis-Inducing Ligand