摘要
利用PCR方法和体内同源重组技术,对耐辐射奇球菌(Deinococcus radiodurans)中控制色素合成的关键基因———crtI进行缺失突变,成功获得红色色素缺失突变株M61。对突变株分别进行不同剂量电离辐射(IR)和不同浓度过氧化氢(H2O2)处理,结果表明:与野生型菌株R1相比,突变株M61对电离辐射的抗性降低;对过氧化氢的敏感性明显上升,在高浓度H2O2条件下表现异常敏感。HPLC分析结果显示,crtI基因的完全缺失对色素合成途径产生重要影响,导致番茄红素和其他红色类胡萝卜素的合成被抑制。证明crtI基因是耐辐射奇球菌中控制红色类胡萝卜素合成的一个关键基因。为阐明耐辐射奇球菌中类胡萝卜素参与的抗辐射和抗氧化机制奠定了一定基础,为进一步研究类胡萝卜素在耐辐射奇球菌中的合成途径及功能提供了思路。
With the method of Polymerase Chain Reaction and homologous genetic recombination in vivo, the key gene encoding bacterial-type phytoene desaturase （CrtI） which controls the carotenoids biosynthesis pathway in the non-photosynthetic and extremely radioresistant bacterium Deinococcus radiodurans was deleted from the genome. The colorless mutant obtained was designated as M61. Survival rates of mutant strain and wild type strain were investigated under different doses of -f-radiation and hydrogen peroxide. The results showed that the radioresistant activity of M61 reduced rapidly under ionization radiation, and it became more sensitive to the treatment of hydrogen peroxide especially to high concentration of hydrogen peroxide compared to that of wild type R1. Reverse Phase High Performance Liquid Chromatography （RP-HPLC） was used to investigate the carotenoid composition of wild type R1 and mutant M61. HPLC results exhibited that the deficient of crtl gene had important effect on pigment biosynthesis pathway, leading to inhibition of the biosynthesis of lycopene and other carotenoids in D. radiodurans. All the results indicated that crtl gene was a key gene controlling the biosynthesis of red carotenoid including lycopene in D. radiodurans. The roles of carotenoids in protecting the bacterial cell from damage by ionization radiation and hydrogen peroxide suggest that the carotenoids contribute to the defense system in D. radiodurans. This study is important for elucidating the radioresistant and antioxidant mechanism in which carotenoids are involved, and it will supply some ideas to the further investigation on the biosynthesis pathway and functions of carotenoids in D. radiodurans.
出处
《微生物学报》
CAS
CSCD
北大核心
2006年第2期210-213,共4页
Acta Microbiologica Sinica
基金
国家"973项目"(2004CB19604)
国家自然科学基金重点资助项目(30330020)
国家杰出青年基金项目(30425038)
国家自然科学基金项目(30500008)~~
