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| Construction and Over-expression of the Activator Gene from Comamonas testosteroni |
| DAI Yi-min;PAN Da-ren;Guangming Xiong;WU Ming-xia;ZHOU Yi-fei |
| College of Crop Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China |
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Abstract The activator gene from Comamonas testosteroni was amplified by PCR from the genomic DNA of C. testosteroni. The PCR products were cloned into plasmid pKtac1 (containing tac promoter), and the recombinant plasmids pKtac1-act1 and pKtac1-act2. were obtained and identified by the analysis of restriction enzymes and DNA sequencing and then respectively transformed into Escherichia coli HB101. The total cell lysate of the host bacteria was extracted to detect the quantity of activator using ELISA. Moreover, the recombinant plasmids were also co-transformed with plasmid pAX1 (containing 3α-HSD/CR gene) into E.coli HB101 respectively. The total cell lysate of the host bacteria was also extracted to detect the quantity of activator and 3α-HSD/CR using ELISA. Moreover, the results indicated that the recombinant plasmids can improve the expression of activator; and the expressions of activator and 3α-HSD/CR can be also improved by plasmid pAX1 co-transformed with recombinant plasmids.
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Received: 21 February 2005
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Corresponding Authors:
ZHOU Yi-fei
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