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Construction and identification of recombinant adenovirus carrying human TIMP-1shRNA gene.
Sun, Y L; Xie, H; Lin, H L; Feng, Q; Liu, Y.
Afiliação
  • Sun YL; The First Hospital of Dalian Medical University, Division of Nephrology, Dalian, China.
  • Xie H; The First Hospital of Dalian Medical University, Division of Nephrology, Dalian, China huaxiecn@126.com.
  • Lin HL; The First Hospital of Dalian Medical University, Division of Nephrology, Dalian, China.
  • Feng Q; The First Hospital of Dalian Medical University, Division of Nephrology, Dalian, China.
  • Liu Y; The First Hospital of Dalian Medical University, Division of Nephrology, Dalian, China.
Genet Mol Res ; 14(1): 199-208, 2015 Jan 16.
Article em En | MEDLINE | ID: mdl-25729951
The aim of this study was to construct the recombinant adenovirus carrying human TIMP-1shRNA gene expression system for preliminary identification to lay the foundation for the further study of gene therapy. Using the Adeno-X system, the recombinant adenovirus plasmid pAdeno-X green fluorescent protein (GFP)-tissue inhibitor of metalloprotease (TIMP)-1 small hairpin (1shRNA) was constructed by including the target gene fragment of the TIMP-1shRNA shuttle plasmid pShuttle2-GFP-TIMP-1shRNA and the backbone plasmid pAdeno-X by homologous recombination in Escherichia coli. Recombinant plasmids were transfected into HEK293A cells to package the recombinant adenovirus rvAdeno-XGFP-TIMP-1shRNA. The recombinant adenovirus was identified by polymerase chain reaction, and the viral titer and infection efficiency were detected using GFP. Polymerase chain reaction and restriction endonuclease digestion demonstrated that rvAdeno-XGFP-TIMP-1shRNA had been successfully constructed, which has a strong ability to infect the kidney. The TIMP-1shRNA adenovirus expression vector was successfully constructed using homologous recombination methods.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Recombinação Genética / Adenoviridae / Inibidor Tecidual de Metaloproteinase-1 / RNA Interferente Pequeno Tipo de estudo: Diagnostic_studies Limite: Animals / Humans Idioma: En Revista: Genet Mol Res Assunto da revista: BIOLOGIA MOLECULAR / GENETICA Ano de publicação: 2015 Tipo de documento: Article País de afiliação: China País de publicação: Brasil
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Recombinação Genética / Adenoviridae / Inibidor Tecidual de Metaloproteinase-1 / RNA Interferente Pequeno Tipo de estudo: Diagnostic_studies Limite: Animals / Humans Idioma: En Revista: Genet Mol Res Assunto da revista: BIOLOGIA MOLECULAR / GENETICA Ano de publicação: 2015 Tipo de documento: Article País de afiliação: China País de publicação: Brasil