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Title: Molecular cloning of ADP-glucose pyrophosphorylase from cyanobacteria

Abstract

Bacterial and higher plant ADP-glucose pyrophosphorylase differ in structure (homotetramer vs heterotetramer respectively) and allosteric activator and inhibitor. However, highly conserved regions can be identified when sequence comparisons are made between ADP-glucose pyrophosphorylases from diverse species. The fructose 1,6 bisphosphate binding site (activator site) in E. coli is highly conserved in all species for which ADP-glucose pyrophosphorylase has been sequenced. A second conserved region, which is labeled by 8-azido-ATP, is also highly conserved in bacteria and higher plants. In previously cloned ADP-glucose pyrophosphorylases the two conserved regions are separated by approximately 80 amino acids. The authors have used these conserved amino acid sequences to design degenerate oligonucleotide primers for polymerase chain reaction amplification (PCR) of part of the ADP-glucose pyrophosphorylase geae. A predicted 240 bp fragment is amplified in PCR reactions using Anabaena sp. PCC 7120 and Synechocystis sp. PCC 6803 genomic DNA as template. The deduced amino acid sequence from the 240 bp Anabaena fragment shares 75 and 76% identity to that of the rice endosperm and spinach leaf ADP-glucose pyrophosphorylases respectively. The Anabaena amino acid sequence shares 42% identity in amino acid sequence to the E. coli enzyme. At the nucleotide level there is 66% identity of themore » Anabaena sequence to rice endosperm ADP-glucose pyrophosphorylase and 54% to the E. coli gene. The PCR amplified fragments are being used to screen respective Anabaena and Synechocystis genomic gene libraries.« less

Authors:
; ; ;  [1]
  1. Michigan State Univ. Plant Research Lab., East Lansing (United States)
Publication Date:
OSTI Identifier:
5403562
Report Number(s):
CONF-9107184-
Journal ID: ISSN 0079-2241; CODEN: PPYSA
Resource Type:
Conference
Journal Name:
Plant Physiology, Supplement; (United States)
Additional Journal Information:
Journal Volume: 96:1; Conference: Annual meeting of the American Society of Plant Physiology, Albuquerque, NM (United States), 28 Jul - 1 Aug 1991; Journal ID: ISSN 0079-2241
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; CYANOBACTERIA; MOLECULAR BIOLOGY; TRANSFERASES; DNA-CLONING; AMINO ACID SEQUENCE; DNA SEQUENCING; GENETICS; BIOLOGY; CLONING; DNA HYBRIDIZATION; ENZYMES; HYBRIDIZATION; MICROORGANISMS; MOLECULAR STRUCTURE; ORGANIC COMPOUNDS; PROTEINS; STRUCTURAL CHEMICAL ANALYSIS; 550200* - Biochemistry

Citation Formats

Kakefuda, G, Charng, Yeeyung, Iglesias, A, and McIntosh, L. Molecular cloning of ADP-glucose pyrophosphorylase from cyanobacteria. United States: N. p., 1991. Web.
Kakefuda, G, Charng, Yeeyung, Iglesias, A, & McIntosh, L. Molecular cloning of ADP-glucose pyrophosphorylase from cyanobacteria. United States.
Kakefuda, G, Charng, Yeeyung, Iglesias, A, and McIntosh, L. 1991. "Molecular cloning of ADP-glucose pyrophosphorylase from cyanobacteria". United States.
@article{osti_5403562,
title = {Molecular cloning of ADP-glucose pyrophosphorylase from cyanobacteria},
author = {Kakefuda, G and Charng, Yeeyung and Iglesias, A and McIntosh, L},
abstractNote = {Bacterial and higher plant ADP-glucose pyrophosphorylase differ in structure (homotetramer vs heterotetramer respectively) and allosteric activator and inhibitor. However, highly conserved regions can be identified when sequence comparisons are made between ADP-glucose pyrophosphorylases from diverse species. The fructose 1,6 bisphosphate binding site (activator site) in E. coli is highly conserved in all species for which ADP-glucose pyrophosphorylase has been sequenced. A second conserved region, which is labeled by 8-azido-ATP, is also highly conserved in bacteria and higher plants. In previously cloned ADP-glucose pyrophosphorylases the two conserved regions are separated by approximately 80 amino acids. The authors have used these conserved amino acid sequences to design degenerate oligonucleotide primers for polymerase chain reaction amplification (PCR) of part of the ADP-glucose pyrophosphorylase geae. A predicted 240 bp fragment is amplified in PCR reactions using Anabaena sp. PCC 7120 and Synechocystis sp. PCC 6803 genomic DNA as template. The deduced amino acid sequence from the 240 bp Anabaena fragment shares 75 and 76% identity to that of the rice endosperm and spinach leaf ADP-glucose pyrophosphorylases respectively. The Anabaena amino acid sequence shares 42% identity in amino acid sequence to the E. coli enzyme. At the nucleotide level there is 66% identity of the Anabaena sequence to rice endosperm ADP-glucose pyrophosphorylase and 54% to the E. coli gene. The PCR amplified fragments are being used to screen respective Anabaena and Synechocystis genomic gene libraries.},
doi = {},
url = {https://www.osti.gov/biblio/5403562}, journal = {Plant Physiology, Supplement; (United States)},
issn = {0079-2241},
number = ,
volume = 96:1,
place = {United States},
year = {1991},
month = {5}
}

Conference:
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