Construction of a recombinant vector based on AAV carrying human endothelial nitric-oxide synthase gene.
Acta Pharmacol Sin
; 24(7): 637-40, 2003 Jul.
Article
de En
| MEDLINE
| ID: mdl-12852827
ABSTRACT
AIM:
To construct an AAV based vector carrying human endothelial nitric-oxide synthase (eNOS) cDNA and study its expression in vitro for future gene therapy.METHODS:
eNOS cDNA was inserted into the EcoR I site of pSNAV-1 containing the cytomegalovirus (CMV) promoter and inverted terminal repeat sequences of adeno-associated virus. The constructed vector was transfected into BHK and C2C12 cells. eNOS cDNA and mRNA were detected by polymerase chain reaction (PCR) and reverse transcription-PCR (RT-PCR), respectively.RESULTS:
By restriction enzyme digestion analysis, it was proved that eNOS cDNA was inserted into pSNAV-1 in a proper direction. PCR detection demonstrated that pSNAV-eNOS was transferred into both BHK and C2C12 cells. RT-PCR analysis showed that these pSNAV-eNOS transfected cells could express eNOS mRNA.CONCLUSION:
pSNAV-eNOS was successfully constructed with the ability to express human eNOS mRNA in cultured mammalian cells.
Recherche sur Google
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Dependovirus
/
Nitric oxide synthase
Limites:
Animals
/
Humans
Langue:
En
Journal:
Acta Pharmacol Sin
Sujet du journal:
FARMACOLOGIA
Année:
2003
Type de document:
Article
Pays d'affiliation:
Chine