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Title: DNA sequence and regulation of the gene (cbpA) encoding the 42-kilodalton cytoplasmic membrane carotenoprotein of the cyanobacterium Synechococcus sp. strain PCC 7942

Abstract

The gene (cbpA) coding for a carotenoid-binding protein of the cyanobacterium Synechococcus sp. strain PCC 7942 (Anacystis nidulans R2) has been cloned and sequenced. A polyclonal antibody against the protein was used to identify immunoreactive clones from a {lambda}gt11 expression library of Synechococcus strain PCC 7942. The initial positive clone ({lambda}gtAN42) contained a 0.9-kilobase (kb) chromosomal fragment, which was used to detect a larger chromosomal fragment from a {lambda}EMBL3 library. The {lambda}EMBL3 recombinant, {lambda}EM109, contained an 18-kb portion of the Synechococcus strain PCC 7942 chromosome. The open reading frame of cbpA encoded 450 amino acids which give rise to a protein of 49,113 daltons. The hydrophobicity plot indicates that the protein may have a 49-residue signal sequence which is cleaved to yield a mature protein of 43,709 daltons. The protein has been localized in the cytoplasmic membrane by biochemical procedures as well as by electron microscopic immunocytochemistry. Northern (RNA) blot analysis indicates that transcription of cbpA is tightly regulated by DNA topology, light intensity, and iron concentration. Transcription is greatly induced by growth under high light intensities and repressed during growth under iron-deficient conditions. The DNA gyrase inhibitor novobiocin specifically inhibited the light-induced transcription. In Northern blots, the gene-specific probemore » hybridized to two size classes of RNA, with lengths of 2.0 and 6.2 kb. Since cbpA appears to be a component of the 6.2-kb transcript, it is likely part of a larger operon.« less

Authors:
; ; ;  [1]
  1. Univ. of Missouri-Columbia (USA)
Publication Date:
OSTI Identifier:
6945890
DOE Contract Number:  
FG02-86ER13516
Resource Type:
Journal Article
Journal Name:
Journal of Bacteriology; (USA)
Additional Journal Information:
Journal Volume: 171:6; Journal ID: ISSN 0021-9193
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; CYANOBACTERIA; GENE REGULATION; PROTEINS; DNA SEQUENCING; CAROTENOIDS; DNA-CLONING; GENE OPERONS; MOLECULAR BIOLOGY; RECOMBINANT DNA; TRANSCRIPTION; CLONING; DNA; DNA HYBRIDIZATION; HYBRIDIZATION; HYDROCARBONS; MICROORGANISMS; NUCLEIC ACIDS; ORGANIC COMPOUNDS; PIGMENTS; STRUCTURAL CHEMICAL ANALYSIS; TERPENES; 550200* - Biochemistry

Citation Formats

Reddy, K J, Masamoto, K, Sherman, D M, and Sherman, L A. DNA sequence and regulation of the gene (cbpA) encoding the 42-kilodalton cytoplasmic membrane carotenoprotein of the cyanobacterium Synechococcus sp. strain PCC 7942. United States: N. p., 1989. Web.
Reddy, K J, Masamoto, K, Sherman, D M, & Sherman, L A. DNA sequence and regulation of the gene (cbpA) encoding the 42-kilodalton cytoplasmic membrane carotenoprotein of the cyanobacterium Synechococcus sp. strain PCC 7942. United States.
Reddy, K J, Masamoto, K, Sherman, D M, and Sherman, L A. 1989. "DNA sequence and regulation of the gene (cbpA) encoding the 42-kilodalton cytoplasmic membrane carotenoprotein of the cyanobacterium Synechococcus sp. strain PCC 7942". United States.
@article{osti_6945890,
title = {DNA sequence and regulation of the gene (cbpA) encoding the 42-kilodalton cytoplasmic membrane carotenoprotein of the cyanobacterium Synechococcus sp. strain PCC 7942},
author = {Reddy, K J and Masamoto, K and Sherman, D M and Sherman, L A},
abstractNote = {The gene (cbpA) coding for a carotenoid-binding protein of the cyanobacterium Synechococcus sp. strain PCC 7942 (Anacystis nidulans R2) has been cloned and sequenced. A polyclonal antibody against the protein was used to identify immunoreactive clones from a {lambda}gt11 expression library of Synechococcus strain PCC 7942. The initial positive clone ({lambda}gtAN42) contained a 0.9-kilobase (kb) chromosomal fragment, which was used to detect a larger chromosomal fragment from a {lambda}EMBL3 library. The {lambda}EMBL3 recombinant, {lambda}EM109, contained an 18-kb portion of the Synechococcus strain PCC 7942 chromosome. The open reading frame of cbpA encoded 450 amino acids which give rise to a protein of 49,113 daltons. The hydrophobicity plot indicates that the protein may have a 49-residue signal sequence which is cleaved to yield a mature protein of 43,709 daltons. The protein has been localized in the cytoplasmic membrane by biochemical procedures as well as by electron microscopic immunocytochemistry. Northern (RNA) blot analysis indicates that transcription of cbpA is tightly regulated by DNA topology, light intensity, and iron concentration. Transcription is greatly induced by growth under high light intensities and repressed during growth under iron-deficient conditions. The DNA gyrase inhibitor novobiocin specifically inhibited the light-induced transcription. In Northern blots, the gene-specific probe hybridized to two size classes of RNA, with lengths of 2.0 and 6.2 kb. Since cbpA appears to be a component of the 6.2-kb transcript, it is likely part of a larger operon.},
doi = {},
url = {https://www.osti.gov/biblio/6945890}, journal = {Journal of Bacteriology; (USA)},
issn = {0021-9193},
number = ,
volume = 171:6,
place = {United States},
year = {1989},
month = {6}
}