【摘要】 旨在鉴定和分析嗜酸氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans)ATCC 23270硫酸盐同化相关的基因。首先利用生物信息学的方法分析A.ferrooxidans ATCC 23270基因组中可能参与硫酸盐同化的基因,通过反转录、凝胶电泳等技术手段对可能参与编码半胱氨酸(Cys)操纵子的几个候选基因cys JI、cys H、cys D-2、cys N、cys D-1和cys NC进行共转录鉴定,并对其关键的基因进行体外克隆、表达、纯化重组,同时进行功能验证和酶活测定。结果分析表明,半胱氨酸(Cys)操纵子的候选基因cys D-1和cys NC共转录,Cys JI、cys H、cys D-2、cys N共转录。初步确定了A.ferrooxidans ATCC 23270硫酸盐同化可能的基本代谢路径,探索了半胱氨酸(Cys)操纵子的调控机制。更多还原

【Abstract】 This work aims to identify and analyze the related genes of sulfate-assimilation in Acidithiobacillus ferrooxidans ATCC 23270. Firstly, the genes involved in sulfate-assimilation in the genome of A. ferrooxidans ATCC 23270 were analyzed using bioinformatics. Then we co-transcripted and identified the candidate genes cys JI, cys H, cys D-2, cys N, cys D-1, and cys NC probably involving in encoding cysteine(Cys)operon by reverse transcription and gel electrophoresis. Further we cloned in vitro, expressed, purified and recombined the key genes, and confirmed their functions and measured their enzyme activities. Results showed that among these candidate genes of Cys operon, cys D-1 and cys NC were co-transcripted, and Cys JI, cys H, cys D-2 and cys N were co-transcripted. The possible basic metabolic pathway of sulfate-assimilation in A. ferrooxidans ATCC 23270 was determined preliminarily, and the regulatory mechanism of Cys operon was explored.更多还原