Construction of an adenovirus vector carrying the human tissue inhibitor of metalloproteinase 2 gene.
Xin Zhao, Hailin Li, Wenbin Ji, Xian-Jie Shi, Jiahong Dong
Abstract
Xin Zhao, Hailin Li, Wenbin Ji, Xian-Jie Shi, Jiahong Dong
Abstract
This study sought to construct an adenoviral vector carrying the human tissue inhibitor of the metalloproteinase 2 (TIMP-2) gene for use in gene therapy. A recombinant adenovirus (AdTIMP-2) containing a human TIMP-2 cDNA fragment was generated by homologous recombination in BJ5183 bacteria. Recombinant plasmids were screened by antibiotic selection. The adenovirus vector was then packaged and amplified in HEK293 cells. A recombinant adenoviral vector carrying human TIMP-2 was constructed. The titer was 4 x 10(11) pfu/mL after purification. The expression of the TIMP-2 gene in HEK293 cells was detected by PCR. A recombinant adenoviral vector carrying human TIMP-2 was successfully constructed and is available for further use in gene therapy for vascular disease.
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This study sought to construct an adenoviral vector carrying the human tissue inhibitor of the metalloproteinase 2 (TIMP-2) gene for use in gene therapy. A recombinant adenovirus (AdTIMP-2) containing a human TIMP-2 cDNA fragment was generated by homologous recombination in BJ5183 bacteria. Recombinant plasmids were screened by antibiotic selection. The adenovirus vector was then packaged and amplified in HEK293 cells. A recombinant adenoviral vector carrying human TIMP-2 was constructed. The titer was 4 x 10(11) pfu/mL after purification. The expression of the TIMP-2 gene in HEK293 cells was detected by PCR. A recombinant adenoviral vector carrying human TIMP-2 was successfully constructed and is available for further use in gene therapy for vascular disease.
Key concepts: Recombinant DNA, Genetic enhancement, HEK 293 cells, Viral vector, Complementary DNA, Plasmid, Molecular biology, Gene