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PMID: 518860 Published · ppublish English Journal Article

Cloning of synthetic deoxyribonucleic acid that codes for embryonic cardiac myosin light-chain polypeptide.

Biochemistry ·Vol. 18 ·No. 25 ·1979-12-11 ·Pages 5641-7

Arnold HH, Siddiqui MA

Abstract

Double-stranded complementary deoxyribonucleic acid (cDNA) transcribed in vitro from a partially pure myosin light-chain messenger ribonucleic acid (mRNA) of the chick embryonic heart was cloned in Escherichia coli strain chi 1776 by using the HindIII cleavage site in the plasmid pBR322. The insertion of essentially full length DNA was achieved by repeated selection of large-size cDNA transcripts. Of the 12 transformants that contained large-size DNA inserts, the clone pML10 insert was 950 base pairs in length, almost the same size as myosin light-chain mRNA (980 nucleosides). The clone pML10 was identified by hybridization with a highly pure cDNA probe and by hybrid-arrested translation assay. pML10 was further characterized by partial restriction enzyme mapping. The availability of a cloned DNA probe for myosin light-chain facilitates the analysis of the mechanism underlying the induction of cardiac muscle specific gene transcription in presumptive heart-forming cells of the chick blastoderm.

MeSH Terms
Animals Chick Embryo Cloning, Molecular DNA/metabolism DNA Restriction Enzymes DNA, Recombinant/metabolism Myocardium/metabolism Myosins/biosynthesis Nucleic Acid Hybridization Plasmids Poly A/metabolism Protein Biosynthesis RNA, Messenger/metabolism Rabbits Reticulocytes/metabolism Transcription, Genetic
Chemicals
DNA, Recombinant RNA, Messenger Poly A DNA DNA Restriction Enzymes Myosins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Arnold H H
Siddiqui M A
Article Info
Journal
Biochemistry
Abbr.
Biochemistry
ISSN
0006-2960
Published
1979-12-11
Pages
5641-7
Language
English
Region
United States
NLM ID
0370623
Subset
IM
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