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PMID: 6089213 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Isolation of cDNA clones for the chicken neural cell adhesion molecule (N-CAM).

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 81 ·No. 17 ·1984-09-00 ·Pages 5584-8

Murray BA, Hemperly JJ, Gallin WJ, MacGregor JS, Edelman GM, Cunningham BA

Abstract

Enriched mRNA coding for the neural cell adhesion molecule (N-CAM) was prepared from 9-day embryonic chicken brains by immunoprecipitation of polysomes with antibodies to N-CAM. This mRNA programmed the translation in vitro of N-CAM polypeptide chains in a rabbit reticulocyte lysate system. Two independent N-CAM cDNA clones (designated pEC001 and pEC020) were derived from the enriched RNA. The specificity of pEC001 for N-CAM mRNA was verified by hybrid selection experiments. Both plasmids hybridized to two discrete 6- to 7-kilobase-long RNA species in poly(A)+ RNA from embryonic chicken brain and to lesser amounts of polydisperse material of smaller sizes (probably degradation products of the large RNAs). No hybridization was detected to poly(A)+ RNA from embryonic liver. Southern blotting experiments with pEC001 detected only one hybridizing fragment in chicken genomic DNA digested with several different restriction enzymes, suggesting that sequences corresponding to those within this region of N-CAM mRNA are present at most only a few times, and possibly only once, in the chicken genome.

MeSH Terms
Animals Antigens, Surface/genetics Base Sequence Brain/metabolism Cell Adhesion Molecules Chick Embryo Cloning, Molecular DNA/isolation & purification DNA Restriction Enzymes Nucleic Acid Hybridization Plasmids Polyribosomes/metabolism Protein Biosynthesis RNA, Messenger/genetics Rats Reticulocytes/metabolism
Chemicals
Antigens, Surface Cell Adhesion Molecules RNA, Messenger DNA DNA Restriction Enzymes
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Murray B A
Hemperly J J
Gallin W J
MacGregor J S
Edelman G M
Cunningham B A
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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.81.17.5584
Published
1984-09-00
Pages
5584-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC391751
Subset
IM
Grants
NICHD NIH HHS · HD-09635 · United States
NICHD NIH HHS · HD-16550 · United States
NCRR NIH HHS · S07 RR07065 · United States
External Links
PubMed source DOI: 10.1073/pnas.81.17.5584 PMC Full Text View journal details
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