title: Expression of human uterine tissue-type plasminogen activator in mouse cells using BPV vectors.
abstract: Human tissue-type plasminogen activator (t-PA) cDNA was cloned from uterine tissue and engineered in expression vectors for production in mouse C127 cells. The vectors consisted of the bovine papilloma virus-1 (BPV-1) genome and t-PA transcriptional unit with a mouse metallothionein (MT-1) promoter at the 5' end and MT-1 genomic sequences or SV40 early introns and polyadenylation signals at the 3' end. Analysis of the expression vectors transfected into cells revealed that t-PA is expressed 100- to 200-fold more with an intronless vector utilizing the SV40 polyadenylation signal than with other, intron-containing vectors. RNA analysis of stable cell lines indicated that t-PA expression levels correlated with mRNA abundance. DNA copy number and transcriptional rate of the MT-1 promoter remained constant in cell lines transformed by different BPV expression vectors. Uterine t-PA produced by recombinant DNA means was enzymatically active and similar in properties to Bowes melanoma t-PA.
authors.affiliation: Integrated Genetics, 31 New York Avenue, Framingham, MA 01701.
authors.forename: V B
authors.initials: VB
authors.lastname: Reddy
authors.forename: A J
authors.initials: AJ
authors.lastname: Garramone
authors.forename: H
authors.initials: H
authors.lastname: Sasak
authors.forename: C M
authors.initials: CM
authors.lastname: Wei
authors.forename: P
authors.initials: P
authors.lastname: Watkins
authors.forename: J
authors.initials: J
authors.lastname: Galli
authors.forename: N
authors.initials: N
authors.lastname: Hsiung
mesh_terms: D000818:Animals; D001483:Base Sequence; D052688:Bovine papillomavirus 1 / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005260:Female; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D007163:Immunosorbent Techniques; D051379:Mice; D008969:Molecular Sequence Data; D027383:Papillomaviridae / Q000235:genetics*; D011061:Poly A / Q000235:genetics; D012326:RNA Splicing; D010959:Tissue Plasminogen Activator / Q000235:genetics* / Q000276:immunology; D014162:Transfection; D014599:Uterus / Q000201:enzymology* / Q000502:physiology
chemical_list: D011061:Poly A; D004247:DNA; D010959:Tissue Plasminogen Activator