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PMID: 6946453 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Transformation of frog embryos with a rabbit beta-globin gene.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 78 ·No. 8 ·1981-08-00 ·Pages 5051-5

Rusconi S, Schaffner W

Abstract

In order to study the fate and possible expression of foreign DNA during embryogenesis of the frog Xenopus laevis, we have injected a rabbit beta-globin gene into fertilized Xenopus eggs. Frog embryo DNA was extracted at various stages of development, fractionated by agarose gel electrophoresis, transferred to nitrocellulose filters, and hybridized to labeled beta-globin recombinant plasmid DNA. It was found that the injected DNA replicated extrachromosomally, reaching, at gastrula stage, a level equivalent to a 10- to 200-fold amplification of input DNA. At later stages, a majority of the foreign DNA was degraded, but a small fraction was maintained. Six-week-old tadpoles as well as six-month-old frogs contained an average of 3-10 copies of the rabbit globin gene per cell. Most of these persisting globin genes were present as long tandem repeats and comigrated in agarose gel electrophoresis with high molecular weight Xenopus DNA. Analysis of globin gene expression by S1 nuclease mapping showed that the rabbit beta-globin promoter was recognized in the frog embryo and that the transcripts were correctly spliced.

MeSH Terms
Animals DNA Replication Extrachromosomal Inheritance Gene Amplification Gene Expression Regulation Genes Globins/genetics Operon Plasmids Rabbits Species Specificity Transcription, Genetic Transformation, Genetic Xenopus laevis/genetics
Chemicals
Globins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Rusconi S
Schaffner W
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40 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.78.8.5051
Published
1981-08-00
Pages
5051-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC320330
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.78.8.5051 PMC Full Text View journal details
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