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PMID: 3883358 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Detection and characterization of a mouse alpha-spectrin cDNA clone by its expression in Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 82 ·No. 5 ·1985-03-00 ·Pages 1367-71

Cioe L, Curtis P

Abstract

A cloned segment of mouse alpha-spectrin mRNA has been identified by immunological techniques. Double-stranded cDNA derived from spleens of anemic mice was introduced into a bacterial expression vector, pUC, and transformed Escherichia coli colonies were screened by using an antiserum to erythrocyte membrane ghost proteins. Of 17 positive colonies, 2 bound antibody to mouse spectrin, and these 2 colonies contained 750-base-pair inserts that cross-hybridized. Transfer of the 750-base-pair insert to an expression vector containing the PL promoter of phage lambda produced larger amounts of peptides that were bound by antibody to mouse spectrin. The spectrin-like peptides made in E. coli elicited antibody that reacted only with the alpha-spectrin subunit of erythrocyte membranes. This clone will be useful for the study of the structure and expression of the spectrin gene, particularly in understanding the role of spectrin in human inherited hemolytic anemias.

MeSH Terms

Animals Antibody Specificity Cloning, Molecular DNA/genetics Erythrocyte Membrane/ultrastructure Escherichia coli/genetics Gene Expression Regulation Mice Peptide Fragments/genetics Spectrin/genetics,immunology

Chemicals

Peptide Fragments Spectrin DNA

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Cioe L

Curtis P

References (26)

26 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.82.5.1367

Published

1985-03-00

Pages

1367-71

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC397262

Subset

Grants

NCI NIH HHS · CA-30136 · United States

External Links

PubMed source DOI: 10.1073/pnas.82.5.1367 PMC Full Text View journal details

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Detection and characterization of a mouse alpha-spectrin cDNA clone by its expression in Escherichia coli.

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