【摘要】 As(Ⅲ)氧化酶能够催化氧化As(Ⅲ),将水体中的As(Ⅲ)氧化成As(Ⅴ)后,毒性将大大降低且易脱除.通过聚合酶链反应(PCR)从能高效氧化As(Ⅲ)的嗜热栖热菌株(Thermus therm ophilusHB8)质粒pTT27扩增了三价砷氧化酶基因(TTHB127,TTHB128),将TTHB127、TTHB128分别定向克隆到pBBR1MCS-5载体上,构建重组质粒127pBBR128,通过三亲杂交,在辅助质粒pRK2013的帮助下,将质粒127pBBR128转移到耐性菌AS-01中,得到的工程菌127pBBR128-AS具有As(Ⅲ)的耐受性和高氧化性的功能,且127pBBR128-AS的As(Ⅲ)氧化酶活力是出发菌株AS-01的8倍.更多还原

【Abstract】 The arsenite oxidase could oxidize arsenite,and oxidation of As(Ⅲ) to As(Ⅴ) could enhance As(Ⅲ) removal and reduce its toxicity.The arsenite oxidase gene fragments TTHB127 and TTHB128 were isolated from the plasmid pTT27 of Thermus thermophilus HB8 by PCR amplification.Recombinant plasmids 127pBBR128 was constructed by ligating TTHB127 and TTHB128 genes into broad host vector pBBR1MCS-5.With the plasmid pRK2013,127pBBR128 was transferred into the strain AS-01of arsenite tolerant to construct 127pBBR128-AS.The construction of genetic engineering bacteria 127pBBR128-AS could tolerate high concentration of arsenite and oxidize it.The relative activity of arsenite oxidase of 127pBBR128-AS was approximately 8 times of AS-01.更多还原