High-level secretion of the four salivary plasminogen activators from the vampire bat Desmodus rotundus by stably transfected baby hamster kidney cells.
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.
Krätzschmar
J
J
Research Laboratories of Schering AG Berlin, Germany.
Haendler
B
B
Bringmann
P
P
Dinter
H
H
Hess
H
H
Donner
P
P
Schleuning
W D
WD
D000818:Animals; D001483:Base Sequence; D002685:Chiroptera / Q000235:genetics*; D006224:Cricetinae; D004247:DNA / Q000235:genetics; D005822:Genetic Vectors; D007668:Kidney / Q000502:physiology; D008969:Molecular Sequence Data; D010957:Plasmids; D010960:Plasminogen Activators / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012471:Salivary Proteins and Peptides / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000235:genetics
D011994:Recombinant Proteins; D012471:Salivary Proteins and Peptides; D004247:DNA; D010960:Plasminogen Activators