|Table of Contents|

Construction and Identification of GGPPS Gene Interference Adenovirus(PDF)

《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

Issue:
2013年02期
Page:
104-107
Research Field:
生命科学
Publishing date:

Info

Title:
Construction and Identification of GGPPS Gene Interference Adenovirus
Author(s):
Zhao Yinjuan1; Lai Shanshan2; Li Chaojun2; Xue Bin2
1.College of Forestry Resources and Environment,Nanjing Forestry University,Jiangsu Key Laboatory for Prevention and Management of Invation Species,Nanjing 210037,China
2.Medical School of Nanjing University,Jiangsu Key Laboratory of Molecular Medicine,Nanjing 210093,China
Keywords:
GGPPS; interference; adenovirus
PACS:
Q29
DOI:
-
Abstract:
To construct the recombinant adenovirus vector expressing small RNA against human GGPPS gene,the interferce primer designed according to GGPPS gene sequence was cloned into pshuttle-H1 vector with Bgl Ⅱ and Hind Ⅲ restriction site.E.coli BJ5183 sensitive bacteria were cotransfected with vector lenealized by Pme Ⅰ enzyme and adenovirus vector pAdEasy-1.The adenovirus was obtained in 293A cells transfected with linearized recombinant adenovirus plasmids.The titer of virus was measured based on the appearing of CPE and was determined by TCID50 assay.The interference efficiency was caculated.The results obtained from DNA sequencing and digestion identification demonstrated that the adenovirus vectors tagerting to human GGPPS gene was constructed.The titer of concentrated virus was 1.995×107 PFU/mL.The interference efficiency of endogenous GGPPS gene in WRL68 cells was above 70% as evidenced by western blot analysis.The interference adenovirus vector of human GGPPS gene is successfully constructed.

References:

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Last Update: 2013-06-30