Journal of Food, Agriculture and Environment
Vol 8,
Issue 3&4,2010
Online ISSN: 1459-0263
Print ISSN: 1459-0255
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Isolation and characterization of the petII promoter of Acidithiobacillus ferrooxidans
Author(s):
Yu Yang,
Zhiying Huang, Mengxue Diao, Guanzhou Qiu *Recieved Date: 2010-07-10, Accepted Date: 2010-11-07
Abstract:A promoter-probe vector (pBST) was constructed that contained lacZ gene of Escherichia coli as a selectable marker gene. Subsequently, potential promoter region of the petII operon from Acidithiobacillus ferrooxidans was cloned into the promoterless pBST vector, and the recombinant was named as pSV72. E. coli DH5α cells carrying pSV72 exhibited approximately 70% β-galactosidease activity compared with those carrying pBST vector, suggesting that the petII promoter was not as strong as the glutamate pyruvate transaminase (gpt) promoter. Furthermore, two putative core elements of the petII promoter, fragments TTGATA (-95 to -90) and TATGCT (-59 to -54), were site-directed mutated to GTGATA (muA) and CATGCT (muB), respectively. The results of a reporter gene assay demonstrated that the two mutations decreased the β-galactosidease activity to 15% and 59% of the wild-type (pSV72), respectively. This suggested that TTGATA and TATGCT were the -35 box and -10 box sequence, respectively, of the petII promoter.
Keywords:Acidithiobacillus ferrooxidans,
petII operon, promoter, promoter-probing vector, site-directed mutagenesisJournal: Journal of Food, Agriculture and Environment
Year: 2010
Volume: 8
Issue: 3&4
Category: Environment
Pages: 1383-1387
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