CLONING OF A GENE ENCODING PROTEIN BELONGING TO ABC TRANSPORTER INVOLVED IN BACTERIAL MAGNETIC PARTICLE SYNTHESIS IN MAGNETOSPIRILLUM MAGNETICUM AMB-1
Magnetospirillum magneticum AMB-1 synthesizes intracellular magnetic particles, magnetite (Fe3 O4 ), enveloped by membrane called magnetosome under micro-aerobic conditions. Initial study of random transposon-based mutagenesis generated 62 nonmagnetic mutants of AMB-1 in a mini-Tn5 library. In or...
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| Format: | Article |
| Language: | English |
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SEAMEO, BIOTROP
2009-06-01
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| Series: | Biotropia: The Southeast Asian Journal of Tropical Biology |
| Online Access: | https://journal.biotrop.org/index.php/biotropia/article/view/62 |
| _version_ | 1797758595327066112 |
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| author | ARIS TRI WAHYUDI |
| author_facet | ARIS TRI WAHYUDI |
| author_sort | ARIS TRI WAHYUDI |
| collection | DOAJ |
| description | Magnetospirillum magneticum AMB-1 synthesizes intracellular magnetic particles,
magnetite (Fe3
O4
), enveloped by membrane called magnetosome under micro-aerobic conditions.
Initial study of random transposon-based mutagenesis generated 62 nonmagnetic mutants of
AMB-1 in a mini-Tn5 library. In order to identify a gene involved in bacterial magnetic particle
(BMP) synthesis in the magnetic bacterium M. magneticum AMB-1, a nonmagnetic mutant from
the library designated as NMA38-4, was analyzed. h e amino acid sequence deduced from the
gene directly interrupted by transposon, ORF4 (1482 bp), showed homology to ATP binding
cassette (ABC) transporter of Mesorhizobium loti with 62 % identity and 74 % similarity. It was
strongly indicated by the occurrence of putative consensus sequence of ATP-binding motifs (ATP-
binding protein). h e ORF4 was subsequently cloned in pET-15b and the recombinant ORF4-
Histag fusion protein was heterologously expressed in Escherichia coli BL21 (DE3) pLysS. A 55 kDa
protein corresponding to the ORF4-Histag fusion protein was obtained after puriï¬ cation using
Ni-NTA column. h is is the ï¬ rst report describing a gene cluster containing gene encoding protein
belonging to ABC transporter organized in an operon which is involved in BMP synthesis.
Key words: Magnetospirillum magneticum AMB-1, Bacterial Magnetic Particle (BMP), ATP
Binding Cassette (ABC) Transporter, transposon mutagenesis. |
| first_indexed | 2024-03-12T18:32:17Z |
| format | Article |
| id | doaj.art-c35a1252368c4540b2070469c5098fee |
| institution | Directory Open Access Journal |
| issn | 0215-6334 1907-770X |
| language | English |
| last_indexed | 2024-03-12T18:32:17Z |
| publishDate | 2009-06-01 |
| publisher | SEAMEO, BIOTROP |
| record_format | Article |
| series | Biotropia: The Southeast Asian Journal of Tropical Biology |
| spelling | doaj.art-c35a1252368c4540b2070469c5098fee2023-08-02T08:17:13ZengSEAMEO, BIOTROPBiotropia: The Southeast Asian Journal of Tropical Biology0215-63341907-770X2009-06-0116110.11598/btb.2009.16.1.62CLONING OF A GENE ENCODING PROTEIN BELONGING TO ABC TRANSPORTER INVOLVED IN BACTERIAL MAGNETIC PARTICLE SYNTHESIS IN MAGNETOSPIRILLUM MAGNETICUM AMB-1ARIS TRI WAHYUDI0Department of Biology, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Bogor, IndonesiaMagnetospirillum magneticum AMB-1 synthesizes intracellular magnetic particles, magnetite (Fe3 O4 ), enveloped by membrane called magnetosome under micro-aerobic conditions. Initial study of random transposon-based mutagenesis generated 62 nonmagnetic mutants of AMB-1 in a mini-Tn5 library. In order to identify a gene involved in bacterial magnetic particle (BMP) synthesis in the magnetic bacterium M. magneticum AMB-1, a nonmagnetic mutant from the library designated as NMA38-4, was analyzed. h e amino acid sequence deduced from the gene directly interrupted by transposon, ORF4 (1482 bp), showed homology to ATP binding cassette (ABC) transporter of Mesorhizobium loti with 62 % identity and 74 % similarity. It was strongly indicated by the occurrence of putative consensus sequence of ATP-binding motifs (ATP- binding protein). h e ORF4 was subsequently cloned in pET-15b and the recombinant ORF4- Histag fusion protein was heterologously expressed in Escherichia coli BL21 (DE3) pLysS. A 55 kDa protein corresponding to the ORF4-Histag fusion protein was obtained after puriï¬ cation using Ni-NTA column. h is is the ï¬ rst report describing a gene cluster containing gene encoding protein belonging to ABC transporter organized in an operon which is involved in BMP synthesis. Key words: Magnetospirillum magneticum AMB-1, Bacterial Magnetic Particle (BMP), ATP Binding Cassette (ABC) Transporter, transposon mutagenesis.https://journal.biotrop.org/index.php/biotropia/article/view/62 |
| spellingShingle | ARIS TRI WAHYUDI CLONING OF A GENE ENCODING PROTEIN BELONGING TO ABC TRANSPORTER INVOLVED IN BACTERIAL MAGNETIC PARTICLE SYNTHESIS IN MAGNETOSPIRILLUM MAGNETICUM AMB-1 Biotropia: The Southeast Asian Journal of Tropical Biology |
| title | CLONING OF A GENE ENCODING PROTEIN BELONGING TO ABC TRANSPORTER INVOLVED IN BACTERIAL MAGNETIC PARTICLE SYNTHESIS IN MAGNETOSPIRILLUM MAGNETICUM AMB-1 |
| title_full | CLONING OF A GENE ENCODING PROTEIN BELONGING TO ABC TRANSPORTER INVOLVED IN BACTERIAL MAGNETIC PARTICLE SYNTHESIS IN MAGNETOSPIRILLUM MAGNETICUM AMB-1 |
| title_fullStr | CLONING OF A GENE ENCODING PROTEIN BELONGING TO ABC TRANSPORTER INVOLVED IN BACTERIAL MAGNETIC PARTICLE SYNTHESIS IN MAGNETOSPIRILLUM MAGNETICUM AMB-1 |
| title_full_unstemmed | CLONING OF A GENE ENCODING PROTEIN BELONGING TO ABC TRANSPORTER INVOLVED IN BACTERIAL MAGNETIC PARTICLE SYNTHESIS IN MAGNETOSPIRILLUM MAGNETICUM AMB-1 |
| title_short | CLONING OF A GENE ENCODING PROTEIN BELONGING TO ABC TRANSPORTER INVOLVED IN BACTERIAL MAGNETIC PARTICLE SYNTHESIS IN MAGNETOSPIRILLUM MAGNETICUM AMB-1 |
| title_sort | cloning of a gene encoding protein belonging to abc transporter involved in bacterial magnetic particle synthesis in magnetospirillum magneticum amb 1 |
| url | https://journal.biotrop.org/index.php/biotropia/article/view/62 |
| work_keys_str_mv | AT aristriwahyudi cloningofageneencodingproteinbelongingtoabctransporterinvolvedinbacterialmagneticparticlesynthesisinmagnetospirillummagneticumamb1 |