Cloning and characterization of a gene, mpsA, encoding a protein associated with intracellular magnetic particles from Magnetospirillum sp. strain AMB-1

Tadashi Matsunaga, Noriyuki Tsujimura, Yoshiko Okamura, Haruko Takeyama

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Proteins located within the lipid bilayer, surrounding the intracellular bacterial magnetic particles (BMP) from Magnetospirillum sp. AMB-1, were separated using SDS-PAGE. Several major proteins of approximate molecular weight 66.2, 35.6, and 24.8 kDa were identified. The N-terminal amino acid sequence of one of these proteins, designated MpsA, was determined and used to design a pair of PCR primers which amplified a 105 bp DNA fragment from AMB-1 genomic DNA. Gene-walking, using anchored PCR, was used to determine the complete nucleotide sequence (954 bp) of the mpsA gene. The mpsA encodes a 317 amino acid protein which does not have an N-terminal cytoplasmic transport signal sequence. Intracellular localization studies were carried out using an mpsA-luc gene fusion expressed in AMB-1 following gene transfer by conjugation. The gene fusion was constructed by cloning a 1.6 kb mpsA fragment upstream of luc in the conjugal plasmid pKLC. The MpsA-Luc fusion protein was preferentially located on the magnetic particle membrane. Although the function of MpsA remains unknown, homology searches suggest similarity with the α subunit of acetyl-CoA carboxylase and the CoA-binding motif. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)932-937
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume268
Issue number3
DOIs
Publication statusPublished - 2000 Feb 24
Externally publishedYes

Fingerprint

Magnetospirillum
Gene encoding
Cloning
Organism Cloning
Genes
Fusion reactions
Gene Fusion
Proteins
Gene transfer
Acetyl-CoA Carboxylase
Amino Acids
Polymerase Chain Reaction
Lipid bilayers
DNA
Lipid Bilayers
Coenzyme A
Protein Sorting Signals
Walking
Polyacrylamide Gel Electrophoresis
Amino Acid Sequence

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

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abstract = "Proteins located within the lipid bilayer, surrounding the intracellular bacterial magnetic particles (BMP) from Magnetospirillum sp. AMB-1, were separated using SDS-PAGE. Several major proteins of approximate molecular weight 66.2, 35.6, and 24.8 kDa were identified. The N-terminal amino acid sequence of one of these proteins, designated MpsA, was determined and used to design a pair of PCR primers which amplified a 105 bp DNA fragment from AMB-1 genomic DNA. Gene-walking, using anchored PCR, was used to determine the complete nucleotide sequence (954 bp) of the mpsA gene. The mpsA encodes a 317 amino acid protein which does not have an N-terminal cytoplasmic transport signal sequence. Intracellular localization studies were carried out using an mpsA-luc gene fusion expressed in AMB-1 following gene transfer by conjugation. The gene fusion was constructed by cloning a 1.6 kb mpsA fragment upstream of luc in the conjugal plasmid pKLC. The MpsA-Luc fusion protein was preferentially located on the magnetic particle membrane. Although the function of MpsA remains unknown, homology searches suggest similarity with the α subunit of acetyl-CoA carboxylase and the CoA-binding motif. (C) 2000 Academic Press.",
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AU - Takeyama, Haruko

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