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PMID: 3099166 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Identification of a nonhistone chromosomal protein associated with heterochromatin in Drosophila melanogaster and its gene.

Molecular and cellular biology ·Vol. 6 ·No. 11 ·1986-11-00 ·Pages 3862-72

James TC, Elgin SC

Abstract

Monoclonal antibodies were prepared against a fraction of nuclear proteins of Drosophila melanogaster identified as tightly binding to DNA. Four of these antibodies were directed against a 19-kilodalton nuclear protein; immunofluorescence staining of the polytene chromosomes localized the antigen to the alpha, beta, and intercalary heterochromatic regions. Screening of a lambda gt11 cDNA expression library with one of the monoclonal antibodies identified a recombinant DNA phage clone that produced a fusion protein immunologically similar to the heterochromatin-associated protein. Polyclonal sera directed against the bacterial lacZ fusion protein recognized the same nuclear protein on Western blots. A full-length cDNA clone was isolated from a lambda gt10 library, and its DNA sequence was obtained. Analysis of the open reading frame revealed an 18,101-dalton protein encoded by this cDNA. Two overlapping genomic DNA clones were isolated from a Charon 4 library of D. melanogaster with the cDNA clone, and a restriction map was obtained. In situ hybridization with these probes indicated that the gene maps to a single chromosome location at 29A on the 2L chromosome. This general strategy should be effective for cloning the genes and identifying the genetic loci of chromosomal proteins which cannot be readily assayed by other means.

MeSH Terms
Amino Acid Sequence Animals Antibodies Base Sequence Chromosomal Proteins, Non-Histone/analysis,genetics Chromosomes/ultrastructure DNA, Recombinant/metabolism Drosophila melanogaster/genetics Electrophoresis, Polyacrylamide Gel Fluorescent Antibody Technique Genes Heterochromatin/ultrastructure
Chemicals
Antibodies Chromosomal Proteins, Non-Histone DNA, Recombinant Heterochromatin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
James T C
Elgin S C
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.6.11.3862-3872.1986
Published
1986-11-00
Pages
3862-72
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC367149
Subset
IM
Grants
NIGMS NIH HHS · GM31532 · United States
Databases
GENBANK
M14131
External Links
PubMed source DOI: 10.1128/mcb.6.11.3862-3872.1986 PMC Full Text View journal details
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