Characterization of Carotenogenesis Genes in the Cyanobacterium Anabaena sp. PCC 7120
Characterization of Carotenogenesis Genes in the Cyanobacterium Anabaena sp. PCC 7120
复制标题
蓝藻鱼腥藻中胡萝卜素生成基因的表征。
DOI：
10.1007/978-1-61779-879-5_18
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Takaichi S
中科院分区：
文献类型：
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作者：
Masukawa H;Mochimaru M;Takaichi S
蓝藻在光合作用中产生多种类胡萝卜素，用于光收集和光保护。目的：阐明紫苏中类胡萝卜素的生物合成途径。pcc7120（也称为nostocsp）。PCC 7120)，我们已经产生了缺乏选定的类胡萝卜素生物合成酶的基因破坏突变体。在这里，我们描述了通过三亲交配来构建突变体。携带目标基因的货物质粒被抗生素抗性盒打断，被转化为toE。含有辅助性质粒的大肠杆菌，通过偶联的方式被引入到水珠胞中。双倒易重组是用质粒上的突变基因代替anabaenenomics中的靶基因。采用高效液相色谱法对所选双重组体中的类胡萝卜素进行鉴定。
Cyanobacteria produce many kinds of carotenoids for light harvesting and light protection in photosynthesis. To elucidate the biosynthetic pathways of carotenoids inAnabaenasp. PCC 7120 (also known asNostocsp. PCC 7120), we have produced gene-disruption mutants lacking selected proposed carotenoid biosynthetic enzymes. Here we describe the construction of mutants by triparental mating. A cargo plasmid, bearing a target gene interrupted by an antibiotic-resistant cassette, is transformed toE. colidonor containing a helper plasmid, and is introduced intoAnabaenacells by conjugation. Double-reciprocal recombination replaces the target genes inAnabaenagenome with mutated ones on the plasmid. Carotenoids in the selected double recombinants are identified using high-performance liquid chromatography.