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Involvement of a Gene Encoding Putative Acetate Kinase in Magnetosome Synthesis in Magnetospirillum magneticum AMB-1

HAYATI Journal of Biosciences · Vol. 13 No. 1 (2009)

DOI
10.4308/hjb.13.1.26
ISSN
2086-4094, 1978-3019
Terbit
2009-12-01
Penerbit
Bogor Agricultural University, Indonesia
Bahasa
eng

Abstrak

A nonmagnetic mutant of Magnetospirillum magneticum AMB-1, designated NMA40, was constructed by mini-Tn5 transposon mutagenesis to identify genes involved in magnetosome synthesis. Transposon delivery was carried out through conjugation between M. magneticum AMB-1 as a recipient and Escherichia coli S17-1 (λ pir) carrying pUTmini-Tn5Km1 as a donor strain. NAM40 did not respond to the magnetic fields and completely lacked of magnetosome in the cell. DNA sequence/gen interrupted by transposon (called flanking DNA) was isolated by inverse PCR and cloned into pGEM-T Easy. Alignment of the DNA sequence of the flanking DNA allowed the isolation of an open reading frame (ORF2) within an operon consisting of three genes. The amino acid sequence deduced from ORF2 showed homology with acetate kinase from Sinorhizobium meliloti (50% identity and 67% similarity), which function for acetate metabolism. Further analysis revealed that upstream of ORF2 is ORF1, had homology with phosphotransacetylase of S. meliloti (67% identity, 77% similarity), and ORF3 located downstream of ORF2, had homology with hypothetical protein of Thermotoga maritima (30% identity, 60% similarity). ORF2 was subsequently isolated, cloned, and overexpressed in Escherichia coli BL21 (DE3) pLysS as an ORF2-Histag fusion polypeptide. Key words: Magnetospirillum magneticum AMB-1, magnetosome synthesis, transposon mutagenesis, cloning, overexpression

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  • IPB University
  • Khairun University
  • Sebelas Maret University
  • Indonesian Center for Agricultural Biotechnology and Genetic Resource Research and Development
  • Sriwijaya University
  • University of Göttingen
  • Universitas Islam Negeri Maulana Malik Ibrahim
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Rekan penulisOrganisasiArtikel bersama
Nisa Rachmania MubarikIPB University5
Rika Indri AstutiIPB University4
Munti YuhanaIPB University3
Antonius SuwantoIPB University3
Anja MeryandiniIPB University3
Suryo WiyonoIPB University2
Yulin LestariIPB University2
Abdul MunifIPB University2
Esti PuspitasariIPB University2
Andreas Adhi SatyaAfiliasi tidak diketahui2
Yepy H. RustamAfiliasi tidak diketahui2
Abdjad Asih NawangsihIPB University2
Waode MunaeniKhairun University1
Widanarni WidanarniIPB University1
Mia SetiawatiIPB University1
Bramantyo Jati PrasojoIPB University1
QatrunnadaIPB University1
Ari SusilowatiSebelas Maret University1
Alina AkhdiyaIndonesian Center for Agricultural Biotechnology and Genetic Resource Research and Development1
Latifah Kosim DarusmanIPB University1
Marini WijayantiSriwijaya University1
Martin EngelhauptUniversity of Göttingen1
Hamim HamimUniversitas Islam Negeri Maulana Malik Ibrahim1
İman RusmanaIPB University1
Yuli Siti FatmaIPB University1
Aris TjahjoleksonoIPB University1
Muhammad Faiz AmriIPB University1
Edi HusenAfiliasi tidak diketahui1
Laksmi AmbarsariIPB University1
Rury Eryna PutriIPB University1
Jun NomuraChiba University1
Uci CahliaIPB University1
Siti SholekhaIPB University1
Akhmad Endang Zainal HasanIPB University1
Sri BudiartiIPB University1
Ni Putu Ratna Ayu KrishantiNational Research and Innovation Agency1
Wulandari Antika PutriIPB University1
Hermawaty AbubakarUniversitas Papua1
Sarah Asih FaulinaAfiliasi tidak diketahui1

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Sumber metadata: OAI-PMH (metadata jurnal) http://journal.ipb.ac.id/index.php/hayati/oai, pertama di-harvest 8 Nov 2026, terakhir terlihat 8 Nov 2026.
Metadata asli (oai_dc) 
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	<dc:title xml:lang="en">Involvement of a Gene Encoding Putative Acetate Kinase in Magnetosome Synthesis in Magnetospirillum magneticum AMB-1</dc:title>
	<dc:creator xml:lang="en">WAHYUDI, ARIS TRI</dc:creator>
	<dc:description xml:lang="en">A nonmagnetic mutant of Magnetospirillum magneticum AMB-1, designated NMA40, was constructed by mini-Tn5 transposon mutagenesis to identify genes involved in magnetosome synthesis. Transposon delivery was carried out through conjugation between M. magneticum AMB-1 as a recipient and Escherichia coli S17-1 (λ pir) carrying pUTmini-Tn5Km1 as a donor strain. NAM40 did not respond to the magnetic fields and completely lacked of magnetosome in the cell. DNA sequence/gen interrupted by transposon (called flanking DNA) was isolated by inverse PCR and cloned into pGEM-T Easy. Alignment of the DNA sequence of the flanking DNA allowed the isolation of an open reading frame (ORF2) within an operon consisting of three genes. The amino acid sequence deduced from ORF2 showed homology with acetate kinase from Sinorhizobium meliloti (50% identity and 67% similarity), which function for acetate metabolism. Further analysis revealed that upstream of ORF2 is ORF1, had homology with phosphotransacetylase of S. meliloti (67% identity, 77% similarity), and ORF3 located downstream of ORF2, had homology with hypothetical protein of Thermotoga maritima (30% identity, 60% similarity). ORF2 was subsequently isolated, cloned, and overexpressed in Escherichia coli BL21 (DE3) pLysS as an ORF2-Histag fusion polypeptide. Key words: Magnetospirillum magneticum AMB-1, magnetosome synthesis, transposon mutagenesis, cloning, overexpression </dc:description>
	<dc:publisher xml:lang="en">Bogor Agricultural University, Indonesia</dc:publisher>
	<dc:date>2009-12-01</dc:date>
	<dc:type>info:eu-repo/semantics/article</dc:type>
	<dc:type>info:eu-repo/semantics/publishedVersion</dc:type>
	<dc:type xml:lang="en">Peer-reviewed Article</dc:type>
	<dc:format>application/pdf</dc:format>
	<dc:identifier>http://journal.ipb.ac.id/hayati/article/view/200</dc:identifier>
	<dc:identifier>10.4308/hjb.13.1.26</dc:identifier>
	<dc:source xml:lang="en">HAYATI Journal of Biosciences; Vol. 13 No. 1 (2006): March 2006; 26</dc:source>
	<dc:source>2086-4094</dc:source>
	<dc:source>1978-3019</dc:source>
	<dc:language>eng</dc:language>
	<dc:relation>http://journal.ipb.ac.id/hayati/article/view/200/66</dc:relation>
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