THE CLONING OF TOLUENE-DEGRADING GENE FROM THE GENOME OF Pseudomonas putida ISOLATE INTO THE Escherichia coli ORIGAMI HOST CELL

Wahjudi, Mariana and lndrawati, Evelin and Suryadjaja, Ernest (2015) THE CLONING OF TOLUENE-DEGRADING GENE FROM THE GENOME OF Pseudomonas putida ISOLATE INTO THE Escherichia coli ORIGAMI HOST CELL. In: International Conference on Biological Science 2015 (ICBS), 18-19 September 2015, Yogyakarta.

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Abstract

One of organic wastes that are often produced by petrochemical industry is toluene, a naturally-slow-decomposed compound. The presence of toluene degrading gene can be useful to accelerate toluene degradation. In this study, we aims to clone the toluene gen from Pseudomonas putida isolated from waste. The toluene degrading, tol-like, gene was derived from P. putida genome cutting with Xbal, BamHI, EcoRI or Sacl. The fragments were then cloned on p UCP 18 into Escherichia coli Origami host cells. Selections of transformants were perfonned on luria-Bertani (LB) agar containing 100 ig/ml of ampicillin and then on Bushnell Haas Agar (BHA) containing toluene as the sole carbon source. There were 31 colonies of E. colitransformants grew on LB-amp plate. Selections on BHA-toluene showed that none of the transformants, E. coli bearing empty plasmid or P. putida could grow on 4, 8 40 or 400 ppm of toluene. It might be that the amounts of carbon are too low at these concentrations. At 1% of toluene, the E. coli bearing empty plasmid and 4 transformants were unable to grow, whilst P. putida grew well. Amongst the 27 transformants able to grow on 1% 8 HAtoluenel 12 transformants contained BamHI-restriction fragment and 9 others contained Xbal-restriction fragment.

Item Type: Conference or Workshop Item (Paper)
Uncontrolled Keywords: toluene, Pseudomonas putida, toluene degrading gene, cloning
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: Faculty of Technobiology > Department of Biology
Depositing User: Eko Setiawan 194014
Date Deposited: 23 Sep 2015 02:55
Last Modified: 25 Apr 2016 08:02
URI: http://repository.ubaya.ac.id/id/eprint/25543

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