Cloning and Overexpression of a Gene Encoding Protein Belong to Tetraricopeptide Involved in Magnetosome Synthesis in Magnetospirillum Magneticum AMB-1  

Oleh:

Wahyudi, Aris Tri

Jenis: Article from Journal - ilmiah nasional
Dalam koleksi: Jurnal Mikrobiologi Indonesia vol. 11 no. 1 (Feb. 2006), page 25.
Topik: tetratricopeptide repeat protein; magnetosome; Magnetospirillum magneticum AMB-1; overexpression; cloning

Ketersediaan

Perpustakaan Pusat (Semanggi) Nomor Panggil: JJ139 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

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Isi artikel

A genomic DNA fragment involved in magnetosome synthesis was isolated from Magnetospirillum magneticum AMB-I through mini-Tn5 transposon mutagenesis. The DNA fragment flanking transposon from the genomic DNA of a non-magnetic mutant, designated as NMA42, was isolated by inverse polymerase chain reaction (inverse PCR), sequenced and aligned against the whole genome sequence of AMB-I. A 6020 bp genomic DNA sequence consisting of four open reading frames (ORFs) organized in an operon was determined. The deduced amino acid sequence of 1977 base pair from ORF4 directly interrupted by transposon homolog with tetratricopeptide repeat domain protein (TPR) from Chlamydophila pneumoniae (30% identity, 46% similarity). The presence of four repeats of a degenerate 34 amino acids consensus sequence indicate that the protein may interact with proteins in the cytoplasm to function during magnetosome synthesis. The ORF4 was subsequently isolated from AMB-1 genome by polymerase chain reaction (PCR), cloned, and overexpressed in Escherichia coil as a 70.8 kDa His-tagged polypeptide.

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