Cloning of genes involved in the biosynthesis of pseudobactin, a high-affinity iron transport agent of a plant growth-promoting Pseudomonas strain.
Cloning of genes involved in the biosynthesis of pseudobactin, a high-affinity iron transport agent of a plant growth-promoting Pseudomonas strain.
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克隆参与假杆菌素生物合成的基因，假杆菌素是植物促进假单胞菌菌株的高亲和力铁转运剂。
DOI：
10.1128/jb.157.1.53-58.1984
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发表时间：
1984
影响因子：
3.2
通讯作者：
Leong,J
中科院分区：
文献类型：
--
作者：
Moores,JC;Magazin,M;Ditta,GS;Leong,J
利用广谱接合粘粒pLAFR 1构建了植物促生假单胞菌B10的DNA基因库。重组粘粒含有平均长度为21.5千碱基对的插入DNA。用N-甲基-N '-硝基-N-亚硝基胍、甲磺酸乙酯或紫外线诱变假单胞菌B_（10）菌株，获得了其黄绿色荧光铁载体（微生物铁转运剂）假杆菌素生物合成缺陷的无荧光突变株。没有黄绿色，荧光突变体缺陷的生产假杆菌素进行了鉴定。非荧光突变体通过与基因库杂交并鉴定荧光transconjugants来单独补充。8个重组cosmetry足以补充154个非荧光突变体。互补的模式表明，至少有12个基因排列在4个基因簇是需要假杆菌素的生物合成。考虑到假杆菌素的结构复杂性，这一最小基因数似乎是合理的。
A gene bank of DNA from plant growth-promoting Pseudomonas sp. strain B10 was constructed using the broad host-range conjugative cosmid pLAFR1. The recombinant cosmids contained insert DNA averaging 21.5 kilobase pairs in length. Nonfluorescent mutants of Pseudomonas sp. strain B10 were obtained by mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine, ethyl methanesulfonate, or UV light and were defective in the biosynthesis of its yellow-green, fluorescent siderophore (microbial iron transport agent) pseudobactin. No yellow-green, fluorescent mutants defective in the production of pseudobactin were identified. Nonfluorescent mutants were individually complemented by mating the gene bank en masse and identifying fluorescent transconjugants. Eight recombinant cosmids were sufficient to complement 154 nonfluorescent mutants. The pattern of complementation suggests that a minimum of 12 genes arranged in four gene clusters is required for the biosynthesis of pseudobactin. This minimum number of genes seems reasonable considering the structural complexity of pseudobactin.