Biodiversitas Journal of Biological Diversity
Vol. 17 No. 1 (2016)

Detection of plasmids and curing analysis in copper resistant bacteria Acinetobacter sp. IrC1, Acinetobacter sp. IrC2, and Cupriavidus sp. IrC4

WAHYU IRAWATI (Department of Biology, Pelita Harapan University. M.H. Thamrin Boulevard 1100, Lippo Karawaci, Tangerang 15811, Banten, Indonesia)
TRIWIBOWO YUWONO (Laboratory of Microbiology, Faculty of Agriculture, Gadjah Mada University. Sleman 55281, Yogyakarta, Indonesia)
AMANDA RUSLI (Department of Biology, Pelita Harapan University. M.H. Thamrin Boulevard 1100, Lippo Karawaci, Tangerang 15811, Banten, Indonesia)



Article Info

Publish Date
01 Jan 2016

Abstract

Irawati W, Yuwono T, Rusli A. 2016. Detection of plasmids and curing analysis in copper resistant bacteria Acinetobacter sp. IrC1, Acinetobacter sp. IrC2, and Cupriavidus sp. IrC4. Biodiversitas 17: 296-300. Acinetobacter sp. IrC1, Acinetobacter sp. IrC2, and Cupriavidus sp. IrC4 were copper resistant bacteria. The aims of the study were to establish correlation between bacterial resistance anthe presence of plasmid and to prove the presence of gene that encodes resistance to copper in plasmid. Plasmid curing was carried out by the addition of ethidium bromide, acridine orange, and SDS in Salt Base Solution broth medium. Detection of copper resistance gene in plasmid was carried out by PCR method using CopA primer. The study showed that plasmid isolation has been successfully performed in Acinetobacter sp. IrC1, Acinetobacter sp. IrC2, and Cupriavidus sp. IrC4. The size of plasmid was approximately more than 21 kb. The most effective curing treatment in Acinetobacter sp. IrC1 was 600-700 µg/ml ethidium bromide that reduced up to three times of copper resistance after curing treatment. Meanwhile, copper resistance in Acinetobacter sp. IrC2 and Cupriadus sp. IrC4 decreased four times after curing treatment using 150-200 µg/mL acridine orange and 3000-3500 µg/mL SDS, respectively. The decrease of copper resistance following plasmid curing treatment suggested that copper resistance gene was encoded by the plasmid. The amplification of CopA gene in the plasmid showed the presence of single band DNA with approximately 1.8 kb. The finding on copper resistance present in the plasmid may open a wider application of bacteria as copper bioremediation agent.

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Journal Info

Abbrev

biodiv

Publisher

Subject

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology

Description

The Biodiversitas Journal was first published in 2000 by the Department of Biology, FMNS, Universitas Sebelas Maret, Surakarta, Indonesia, then in 2006 it was co-published by the Society for Indonesian Biodiversity and that department; since 2017 it was also hosted by Smujo. From 2003-2012 it was ...