Cloning ofGenomicDNA Fragment Involved in Acid-Aluminium Tolerancein Bradyrhizobiumjaponicum 38 Through Transposon Mutagenesis  

Oleh:

Wahyudi, Aris Tri ; Mubarik, Nisa Rachmania ; Astuti, Rika Indri

Jenis: Article from Journal - ilmiah nasional
Dalam koleksi: Jurnal Mikrobiologi Indonesia vol. 11 no. 1 (Feb. 2006), page 35.
Topik: cloning; transposon mutagenesis; acid-aluminium tolerance; Bradyrhizobiumjaponicum

Ketersediaan

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

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Isi artikel

An acid-aluminium sensitive mutant of Bradyrhizobium japonicum 38, designated as AAS38, was generated by mini-Tn5 transposon mutagenesis. The experiment was carried out to identify acid-aluminium tolerance gene (AAT) in B. japonicum. Transposon delivery was carried out through conjugation between Escherichia coli S17-1 ~ pir) carrying pUTmini-Tn5Kml and acid-Al tolerant B. japonicum with different of mating time. Frequency of transconjugation was in the range of l0~'-10~. A mutant AAS38 was not able to grow on the Ayanaba medium (pH 4.5) containing 50 ~sMaluminium. However, this mutant formed root nodule of soybean and Siratro plants indicating thegene involved in acid-Al tolerance was not related with nodulation. A 0.8 kb of the genomic DNA fragment flanking the transposon involved in acid-aluminium tolerance was successfully isolated by inverse polymerase chain reaction(Inverse PCR) from AAS38 genome. This fragment was subsequently cloned into pGEM-T Easy (-3 kb) to yield a recombinant plasmid, designated as pGEMT-38 (-3.8 kb), and sequenced. DNA sequence analysis revealed that the genomic DNA fragment had high homology to inner membrane protein from Salmonella typhimurium (80% identityand 86% similarity, E-value= 8xe-' 2) predicted function as efflux transporter.

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