Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: DrugBank:EXPT01586 (G418)
 2,237 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A retrovirus-mediated transduction of B-domain-deleted human blood coagulation factor VIII (FVIII-B) was attempted in partially hepatectomized rats. FVIII-B cDNA was inserted into a retroviral vector (pLNSX) and infective recombinant virus particles were produced in packaging cell lines (psi2 and PA317). Transfection of mouse NIH-3T3 cells with the FVIII-B cDNA inserted recombinant viruses, followed by G418 selection, gave a viral titer of 3.5 x 10(4) CFU/ml. FVIII-B protein, as well as FVIII-B mRNA, was detected in these cells. Transfusion of FVIII-B-expressing retrovirus particles into the tail vein of rats subjected to partial hepatectomy resulted in a relatively higher level of FVIII-B expression in liver and circulating plasma as compared with the sham-operated rats. These results indicate that the augmentation of FVIII activity in the blood of an animal by retroviral gene delivery can be enhanced by partial hepatectomy, and that the retrovirus-mediated FVIII-B cDNA delivery to regenerating liver may be an alternative method for the expression of FVIII-B cDNA in vivo.
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PMID:An efficient retrovirus-mediated transduction of human blood coagulation factor VIII cDNA in regenerating rat liver. 1039 Nov 1

To investigate the non-viral vector mediating human coagulation factor VIII gene expression in mouse 32D cell line, a recombinant plasmid vector, pRC/RSV-hFVIIIBDcDNA, was constructed by cloning B-domain-deleted (Delta760aa-1639aa) human factor VIII cDNA (hFVIIIBDcDNA) into plasmid vector, pRC/RSV. The plasmid RC/RSV-hFVIIIBDcDNA was then transfected by means of SuperFect Transfection Reagent into mouse 32D cell line. After screening with G418, the procoagulant activity (hFVIII:C) and antigen (hFVIII:Ag) of human factor VIII in the culture medium were detected using one-stage method and ELISA, respectively. Furthermore, RT-PCR was performed to observe the transcription of hFVIIIBDcDNA. The results showed that human coagulation factor VIII protein existed in culture medium with hFVIII:C up to 2.01 U/(10(6) cell x 24 hours) and hFVIII:Ag to 450.08 ng/(10(6) cell x 24 hours). RT-PCR displayed mRNA of hFVIIIBDcDNA in 32D cells. It is concluded that the recombinant plasmid RC/RSV-hFVIIIBDcDNA can successfully express human FVIII in mouse 32D cell line, and hFVIII expressed in vitro presents the similar coagulant activity to the native hFVIII existing in normal human plasma.
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PMID:[Non-viral vector mediating human coagulation factor VIII gene expression in mouse 32D cell line]. 1563 47