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Biotechnology
  Year: 2017 | Volume: 16 | Issue: 4-6 | Page No.: 165-173
DOI: 10.3923/biotech.2017.165.173
Characterization of a Naphthalene Catabolic Gene Cluster and Heterologous Expression of Naphthalene Dioxygenase Genes from Rhodococcus ruber OA1
Ying Sun, Zhenglong Wang, Shuyao Zhang, Xiaodan Li, Huaxin Gao and Chunyang Zhang

Abstract:
Background and Objective: Naphthalene is a Polycyclic Aromatic Hydrocarbon (PAH) pollutant which is toxic and widespread in the environment. Rhodococcus ruber OA1 is a gram-positive bacterium which can utilize naphthalene as the sole carbon and energy source for growth, but no molecular biological research has been conducted on its naphthalene catabolic gene cluster. The objective of this study was to characterize the naphthalene catabolic gene cluster and naphthalene dioxygenase (NDO) from R. ruber OA1. Methodology: The gene cluster for naphthalene degradation from R. ruber OA1 was identified by in situ hybridization and sequence analysis. Then the genes narAa and narAb encoding the large and small subunit of naphthalene dioxygenase, as part of the cluster were sub-cloned and heterologously expressed to confirm their functions. Results: In situ hybridization and sequence analysis revealed a 43,754 bp fragment in the cloned plasmid, including the gene cluster narAaAbBC encoding the proteins responsible for converting naphthalene to salicylaldehyde. After the narAa and narAb genes were sub-cloned and expressed, the enzymatic activity was assayed, which revealed that narAa and narAb were functional in Escherichia coli BL21 (DE3). Sub-cloning and heterologous expression of the rub1 gene revealed that the rub1 gene product was not indispensable for naphthalene degradation. Conclusion: A gene cluster for naphthalene biodegradation was identified in R. ruber OA1 for the first time. The narAa and narAb genes were sub-cloned and heterologously expressed to confirm their functions.
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How to cite this article:

Ying Sun, Zhenglong Wang, Shuyao Zhang, Xiaodan Li, Huaxin Gao and Chunyang Zhang, 2017. Characterization of a Naphthalene Catabolic Gene Cluster and Heterologous Expression of Naphthalene Dioxygenase Genes from Rhodococcus ruber OA1. Biotechnology, 16: 165-173.

DOI: 10.3923/biotech.2017.165.173

URL: https://scialert.net/abstract/?doi=biotech.2017.165.173

 
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