使用59-碱基(59be)保守序列和整合酶基因的引物进行PCR扩增,研究了焦化废水处理装置中微生物菌群的基因盒结构,并对两株分离的苯酚降解菌IS—17和IS-46的基因盒进行了扩增测序分析.从5个焦化废水生物膜总DNA样品中均获得了基因盒扩增产物.对IS-17和IS-46的基因盒PCR产物克隆建库并测序,通过序列比对等方法来研究和分析序列的功能.发现本研究获得的基因盒与已报道的基因盒在读码框序列、两端引物结合序列、间隔区大小等方面存在一些不同.本研究结果表明,在焦化废水生物膜菌群中普遍存在基因盒结构,采用基因盒PcR能够从环境或菌株中获取新基因.
To investigate the structure of integron-gene cassette system of the microorganisms in coking wastewater and analyze the gene cassette sequences of two strains of phenol degrading bacteria isolated from identical activated sludge, the gene cassette PCR was performed using the primers designed according to the conserved sequence of the 59 base element structure and the integrase encoding sequence from class 1 integrons. The result showed that all the biofilm samples in the coking wastewater and isolated bacteria produced positive bands. The clones derived from the gene cassette clone libraries of two bacterial strains Alcaligenesfaecalis IS-17 and IS-46 were sequenced and further analyzed by sequence alignment and prediction of their functions. The structures of the gene cassettes were compared with the ones previously reported, in the boundary sites for primer binding, length of region between the ORF and 59be conserved sequence. It was concluded that the gene cassettes structure is widespread in the microbial communities of the coking wastewater biofilm. This work confirms that the gene cassette PCR is an effective method to reveal novel genes from the environment or from the microorganisms.