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PMID: 375197 Published · ppublish English Journal Article

Cloning, mapping and expression of the genetic determinant that encodes for the K88ab antigen.

Nucleic acids research ·Vol. 6 ·No. 3 ·1979-03-00 ·Pages 849-65

Mooi FR, de Graaf FK, van Embden JD

Abstract

The K88 antigen, a plasmid-specified virulence factor of E. coli involved in porcine neonatal diarrhoea, is often found to be associated with the ability to metabolize raffinose (Raf). Plasmid pRI8801 (51 megadalton) was used to clone the determinants of K88 and Raf with the vector pBR322. K88 was found to be encoded by a 7.7 megadalton HindIII fragment. The expression was highly dependent on the orientation of the HindIII fragment within pBR322. By in vitro generation of deletions, the HindIII fragment was reduced in size to 4.3 megadalton. The expression of K88 by pRI8801 and the recombinant plasmids was studied using an enzyme-linked immunosorbent assay. Raf was found to be located on a 4.0 megadalton SalI fragment. A physical map of pRI8801 was constructed. The K88 antigen and Raf genes are not closely linked but separated by a stretch of DNA of about 20 megadalton.

MeSH Terms
Antigens, Bacterial DNA Restriction Enzymes DNA, Recombinant/metabolism Escherichia coli/immunology,metabolism Molecular Weight Plasmids/drug effects Protein Biosynthesis Tetracycline/pharmacology Transcription, Genetic
Chemicals
Antigens, Bacterial DNA, Recombinant DNA Restriction Enzymes Tetracycline
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Mooi F R
de Graaf F K
van Embden J D
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27 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/6.3.849
Published
1979-03-00
Pages
849-65
Language
English
Region
England
NLM ID
0411011
PMCID
PMC327738
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/6.3.849 PMC Full Text View journal details
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