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PMID: 6100967 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Bovine papilloma virus deoxyribonucleic acid: a novel eucaryotic cloning vector.

Molecular and cellular biology ·Vol. 1 ·No. 6 ·1981-06-00 ·Pages 486-96

Sarver N, Gruss P, Law MF, Khoury G, Howley PM

Abstract

A novel eucaryotic vector derived from the transforming region of bovine papilloma virus was established and demonstrated to be highly effective for introducing foreign genes into animal cells. The foreign deoxyribonucleic acid (DNA) is replicated and actively transcribed as an episome, and the transcripts are translated into an authentic gene product. We have constructed a DNA hybrid molecule, BPV69T-rI1, containing the transforming region of bovine papilloma virus DNA and the rat preproinsulin gene I (rI1), and used it to transform susceptible mouse cells. DNA hybridization analysis has demonstrated the presence of multiple unintegrated copies of hybrid DNA molecules, with the bovine papilloma virus 1 DNA segment and the rI1 gene covalently linked in selected transformed cell lines. S1 nuclease analysis revealed the presence of a correctly spliced coding segment of the preproinsulin transcript similar or identical in its electrophoretic mobility to that of messenger ribonucleic acid produced in rat insulinoma cells. Significant levels of a protein immunoreactive with anti-insulin serum were detected by radioimmunoassay in the culture medium of transformed cells. Immunoprecipitation analysis in conjunction with competitive binding to bovine proinsulin established the identity of the protein as that of rat proinsulin.

MeSH Terms
Animals Bovine papillomavirus 1/genetics Cell Line, Transformed DNA, Viral/genetics Genetic Vectors Insulin Papillomaviridae/genetics Proinsulin/biosynthesis,genetics Protein Precursors/genetics RNA, Viral/genetics
Chemicals
DNA, Viral Insulin Protein Precursors RNA, Viral preproinsulin Proinsulin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Sarver N
Laboratory of Pathology, National Cancer Institute, Bethesda, Maryland 20205.
Gruss P
Law M F
Khoury G
Howley P M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.1.6.486-496.1981
Published
1981-06-00
Pages
486-96
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC369692
Subset
IM
Grants
NCI NIH HHS · 1 F32 CA06684-01 · United States
External Links
PubMed source DOI: 10.1128/mcb.1.6.486-496.1981 PMC Full Text View journal details
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