dc.contributor.author | Wahyudi, Aris Tri | |
dc.date.accessioned | 2011-03-22T07:31:40Z | |
dc.date.available | 2011-03-22T07:31:40Z | |
dc.date.issued | 2007 | |
dc.identifier.issn | 1978-3477 | |
dc.identifier.uri | http://repository.ipb.ac.id/handle/123456789/42827 | |
dc.description.abstract | A rapid and simple method to amplify genomic DNA sequences flanking mini-Tn5 transposon insertion was developed. This technique can be used to determine the location and orientation of the transposon insertion within genomic DNA of the bacteria. Based on the mini-Tn5Km1 transposon sequence, PCR primers can be designed to specifically amplify the DNA sequences flanking mini-Tn5 transposon by inverse polymerase chain reaction (inverse PCR) directly, upstream and downstream of the transposon insertion. The method involves: (i) digestion with a restriction enzyme that does not cut mini-Tn5Km1 sequence; (ii) self-ligation under conditions favoring the production of monomeric circles; and (iii) inverse PCR reaction using primers designed from mini- Tn5Km1 sequence to amplify the DNA sequences flanking mini-Tn5Km1 transposon insertion. Feasibility and reliability of this method were demonstrated with mini-Tn5Km1 mutants of the microaerobic magnetic bacterium Magnetospirillum magneticum AMB-1 which are defective in magnetosomes synthesis. The inverse PCR products amplified from these mutant genomes showed the correct fragments as determined through Southern hybridization and DNA sequence analysis. | en |
dc.publisher | IPB (Bogor Agricultural University) | |
dc.relation.ispartofseries | Vol.1;No.1 | |
dc.title | Rapid and Simple Amplification of Genomic DNA Sequences Flanking Transposon | en |
dc.title.alternative | Microbiology Indonesia Vol.1 No.1 Tahun 2007 | en |