|Table of Contents|

Construction and Identification of cnksr2 Gene Interference Adenovirus(PDF)

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

Issue:
2013年03期
Page:
103-107
Research Field:
生命科学
Publishing date:

Info

Title:
Construction and Identification of cnksr2 Gene Interference Adenovirus
Author(s):
Liang Jin1Shen Ning1Xu Kang2Li Chaojun1Xue Bin1
(1.Medical School of Nanjing University,Jiangsu Key Laboratory for Molecular Medicine,Nanjing 210093,China) (2.School of Life Sciences,Nanjing Normal University,Jiangsu Key Laboratory for Molecular and Medical Biotechnology,Nanjing 210023,China)
Keywords:
cnksr2interferenceadenovirus
PACS:
Q291
DOI:
-
Abstract:
To construct a recombinant adenovirus vector expressing small RNA against mouse cnksr2 gene,the interferce primer were designed according to cnksr2 gene sequence,which was cloned into pshuttle-H1 vector with Bgl Ⅱ and Hind Ⅲ restriction site.E.coli BJ5183 sensitive bacterias were cotransfected with lined vector cutted by Pme I enzyme and adenovirus vector pAdEasy-1.The obtained recombinant adenovirus vector was cutted with Pac Ⅰ enzyme,then adenovirus was obtained in 293A cells transfected with lined 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 recombinant adenovirus vector targeting to cnksr2 gene was constructed as evidenced by DNA sequencing and identification of enzyme digestion,which titer of concentrated virus was 3.98×107 PFU/mL.The expression of endogenous cnksr2 mRNA in PC12 cells was specificially shutdown by the recombinant adenovirus vector with an efficiency above 50%.The interference of cnksr2 gene was successfully constructed.

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Last Update: 2013-09-30