Construction of an inducible nitric-oxide synthase gene transferring vector mediated by retrovirus

Meng Wei Zang, Xue Xian Peng, Ping Hu, Jing Sheng Liu

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

AIM: To construct an inducible nitric-oxide synthase (iNOS) gene transferring vector mediated by retrovirus. METHODS: Recombinant DNA and polymerase chain reaction (PCR) amplification techniques were used. RESULTS: With 2 steps of molecular cloning, the full-length cDNA encoding macrophage iNOS was isolated from plasmid pKSiNOS and subcloned into intermediate vector pSP72, adjusting the restriction enzyme sites in both 5'- and 3'-flanking ends of insert fragment. The retroviral vector pLNCXiNOS which contains iNOS coding region, cytomegalovirus promoter and neomycin resistance (neo(r)) gene was further constructed. The authenticity of insertion size and orientation of iNOS sequence was verified by restriction mapping and PCR analysis with iNOS gene-specific primers. CONCLUSION: Retroviral expression vector carrying iNOS fragment is obtained, which provides a material to establish a model of iNOS gene-modified neurons.

Original languageEnglish (US)
Pages (from-to)121-127
Number of pages7
JournalActa Pharmacologica Sinica
Volume19
Issue number2
StatePublished - Mar 31 1998
Externally publishedYes

Keywords

  • Complementary DNA
  • Eukaryotic cells
  • Gene expression regulation
  • Genetic vectors
  • Nitric-oxide synthase
  • Polymerase chain reaction
  • Recombinant DNA
  • Restriction mapping
  • Retroviridae

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)

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