High-level secretion of the four salivary plasminogen activators from the vampire bat Desmodus rotundus by stably transfected baby hamster kidney cells

Gene. 1992 Jul 15;116(2):281-4. doi: 10.1016/0378-1119(92)90526-u.

Abstract

The cDNAs coding for the four Desmodus rotundus salivary plasminogen activators (DSPAs) were subcloned into the mammalian expression vector, pMPSV/CMV, which carries the myeloproliferative sarcoma virus promoter and the cytomegalovirus enhancer. These constructs were transfected, together with plasmids harbouring Geneticin (G418)-resistance and puromycin-resistance genes, into baby hamster kidney cells. Through the selective pressure of both antibiotics, cell clones constitutively overexpressing the DSPA alpha 1, DSPA alpha 2, DSPA beta or DSPA gamma cDNAs were obtained. Secretion of active DSPAs was confirmed by zymographic analysis and quantified using a fibrin plate assay and ELISA.

MeSH terms

  • Animals
  • Base Sequence
  • Chiroptera / genetics*
  • Cricetinae
  • DNA / genetics
  • Genetic Vectors
  • Kidney / physiology
  • Molecular Sequence Data
  • Plasmids
  • Plasminogen Activators / genetics*
  • Plasminogen Activators / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Salivary Proteins and Peptides / genetics*
  • Salivary Proteins and Peptides / metabolism
  • Transfection / genetics

Substances

  • Recombinant Proteins
  • Salivary Proteins and Peptides
  • DNA
  • Plasminogen Activators