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| Construction of the plasmid vector of teiR gene from Comamonas testosteroni and overexpression in E. coli |
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| 350881198110170595 |
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Abstract The teiR gene was amplified by PCR from the genomic DNA of C. testosteroni. The PCR products were cloned into plasmid pKtac2 (containing tac promoter)and pK18, constructing the recombinant plasmids pKtac2-teiR and pKteiR100.The two plasmids were transformed into Escherichia coli HB101 respectively. The total cell lysate of the host bacteria was extracted to detect the quantity of teiR using ELISA. The results showed that plasmid containing tac promoter (pKtac2-teiR) had a higher transcription level than plasmid pKteiR100. Meanwhile, the recombinant plasmids were respectively co-transformed into E. coli HB101 with plasmid p6 (containing 3a-HSD/CR gene).The total cell lysate of the host bacteria was extracted to detect the quantity of teiR and 3a-HSD/CR using ELISA. The results indicated that teiR could significantly promote the 3a-HSD/CR gene expression; and the teiR expression of pKtac2-teiR was higher than that of pKteiR100 after co-transformation with p6. But interestingly , the expression quantity of 3a-HSD/CR co-transformed with plasmids pKtac2-teiR and p6 was less than that co-transformed with plasmids pKteiR100 and p6.
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Received: 27 November 2007
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