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Issue 03
Zhi-Wei Chen#, Xiu-Juan Dong#, Rui-Shen Xie, Xu-yin Yang, Long-Wang Zhang, Long-Hui Zhou, Feng-Mei Yuan, Hang-Ping Wang, Shu-Hua Song*, Jian Liu*
Ibrain 2015;1(3):75-83 Download
Abstract
Background/aim: To construct recombinant vector of over expression and siRNA of Homo sapiens dihydropyrimidine 2 (Dpysl2) gene. Methods: Fragment containing Dpysl2 ORF gene or Dpysl2-siRNA was inserted into recombinant plasmid HIV, then the recombinant of over expression vector was identified by digestion with Xhol and EcoR1, and sequencing of ORF cloning. Pseudovirion containing the recombinants were produced by virus packaging with 293Tα cell, and then the pseudovirion was transfected into HT1299 cell to detect the virus titer.  PC12 were divided into two groups: transfected pseudo virus particles and the control. RT-PCR and WB analysis were performed to observe the changes of Dpysl2 expression after transfection. Results: Successful recombinant vector of over expression and siRNA were identified by digestion with Xhol and EcoR1 and sequencing results. At 48h post-transfection, both of the mRNA and protein level of PC12 were up-regulated in transfected group compared to control group, P<0.05 and P<0.01, respectively. Conclusion: Recombinant vectors of Dpysl2 could effectively regulate the expression of Dpysl2.
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