琼氏不动杆菌GXP04的苯酚羟化酶基因的克隆及序列分析  

Cloning and sequencing of phenol hydroxylase gene from Acinetobacter junii GXP04

作　　者：唐莉丽[1] 孙栋[1] 武波[1] 

机构地区：[1]广西大学生命科学与技术学院微生物及植物遗传工程教育部重点实验室

出　　处：《广西农业生物科学》2006年第1期18-23,共6页Journal of Guangxi Agricultural and Biological Science

基　　金：国家863计划资助项目(2003AA214040)

摘　　要：提取琼氏不动杆菌GXP 04的总DNA,采用限制性内切酶E coRⅠ酶切处理后,构建以pLAFR 3为载体的基因组文库。通过TA IL-PCR扩增出苯酚羟化酶的上游基因,对菌落原位杂交获得的11个转化子进一步鉴定,最终确定其中的8个转化子含有完整的苯酚羟化酶基因,选取其中的pLAFR 3-7对GXP 04的苯酚羟化酶基因的核苷酸序列进行分析,并对其蛋白质基本性质进行预测。Acinetobacter junii GXP04 is a phenol-degrading bacterium isolated before. DNA of GXP04 was digested with EcoR Ⅰ , and ligated with cosmid vector pLAFR3 to construct a genomic library. Colonies were screened by bolt hybridization, and 11 positive clones were selected. According to TAIL-PCR, the upstream gene of phenol hydroxylase was gained and was used to identify the credibility of the 11 positive clones. As a result, only 8 positive clones had the complete phenol hydroxylase. The pLAFR3-7 was selected for the analysis of nucleotide sequence and the basic character of protein.

关 键 词：琼氏不动杆菌GXP04 苯酚 基因组文库 苯酚羟化酶 

分 类 号：X172[环境科学与工程—环境科学] Q78[生物学—分子生物学]