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首页> 外文期刊>Journal of bacteriology >Cloning and sequencing of a 2,5-dichloro-2,5-cyclohexadiene-1,4-diol dehydrogenase gene involved in the degradation of gamma-hexachlorocyclohexane in Pseudomonas paucimobilis.
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Cloning and sequencing of a 2,5-dichloro-2,5-cyclohexadiene-1,4-diol dehydrogenase gene involved in the degradation of gamma-hexachlorocyclohexane in Pseudomonas paucimobilis.

机译：2，5-二氯-2，5-环己二烯-1，4-二醇脱氢酶基因的克隆和测序，涉及革氏假单胞菌中γ-六氯环己烷的降解。

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In Pseudomonas paucimobilis UT26, gamma-hexachlorocyclohexane (gamma-HCH) is converted to 2,5-dichloro-2,5-cyclohexadiene-1,4-diol (2,5-DDOL), which is then metabolized to 2,5-dichlorohydroquinone. Here, we isolated from the genomic library of UT26 two genes which expressed 2,5-DDOL dehydrogenase activity when they were transformed into P. putida and Escherichia coli. Both gene products had an apparent molecular size of 28 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The first gene, named linC, located separately from the two genes (linA and linB) which we had already cloned as genes involved in the gamma-HCH degradation. The other, named linX, located about 1 kb upstream of the linA gene encoding gamma-HCH dehydrochlorinase. A gamma-HCH degradation-negative mutant, named UT72, which lacked the whole linC gene but had the intact linX gene was isolated. The linC gene given in a plasmid could complement UT72. These results strongly suggest that the linC gene but not the linX gene is essential for the assimilation of gamma-HCH in UT26. Deduced amino acid sequences of LinC and LinX show homology to those of members of the short-chain alcohol dehydrogenase family.
机译：在古铜假单胞菌UT26中，γ-六氯环己烷（γ-HCH）被转化为2,5-二氯-2,5-环己二烯-1,4-二醇（2,5-DDOL），然后被代谢为2,5-二氯氢醌。在这里，我们从UT26的基因组库中分离了两个基因，这些基因在转化为恶臭假单胞菌和大肠杆菌后均具有2,5-DDOL脱氢酶的活性。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，两种基因产物的表观分子大小均为28 kDa。第一个名为linC的基因与我们已经作为与丙型六氯环己烷降解有关的基因而克隆的两个基因（linA和linB）分开定位。另一个名为linX，位于编码γ-六氯环己烷脱氢酶的linA基因上游约1 kb。分离出一个γ-HCH降解阴性突变体，名为UT72，该突变体缺少完整的linC基因，但具有完整的linX基因。质粒中给出的linC基因可以补充UT72。这些结果强烈表明，linC基因而非linX基因对于UT26中的γ-六氯环己烷同化至关重要。推导的LinC和LinX的氨基酸序列显示出与短链醇脱氢酶家族成员的同源性。

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