Site directed chromosomal marking of a fluorescent pseudomonad isolated from the phytosphere of sugar beet; Stability and potential for marker gene transfer
Site directed chromosomal marking of a fluorescent pseudomonad isolated from the phytosphere of sugar beet; Stability and potential for marker gene transfer
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DOI：
10.1111/j.1365-294x.1995.tb00276.x
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发表时间：
1995-12-01
影响因子：
4.9
通讯作者：
Ellis, RJ
中科院分区：
文献类型：
--
作者：
Bailey, MJ;Lilley, AK;Ellis, RJ
从甜菜微生物区系中筛选出一株无质粒、非致病的核糖体RNA1组荧光假单胞菌SBW25，并对其进行了改造，使其含有结构性表达的标记基因。通过定点同源重组，将KX盒[卡那霉素抗性(Kan(R))和邻苯二酚2，3-双加氧酶(XylE)]和ZY盒[乳糖利用(lacZY、β-半乳糖苷酶、乳糖渗透酶)]分别引入到细菌的6.6MbP染色体上。选择不同的地点提供灵敏的检测方法，并评估转基因微生物SBW25EeZYGKX在种子接种后在甜菜植株的叶和根上定殖时的标记基因稳定性。
A plasmid-free, non-pathogenic, ribosomal RNA group 1 fluorescent pseudomonad, Pseudomonas fluorescens SBW25, was selected from the microflora of sugar beet (Beta vulgaris) and modified to contain constitutively expressed marker genes. By site directed homologous recombination a KX cassette [kanamycin resistance (kan(r)) and catechol 2,3 dioxygenase (xylE)] and a ZY cassette [lactose utilization (lacZY, beta-galactosidase, lactose permease)] were introduced at least 1 Mbp apart on the 6.6 Mbp bacterial chromosome. Separate sites were selected to provide sensitive detection methods and allow assessments of marker gene stability of the genetically modified micro-organism (GMM), SBW25EeZYGKX, when it colonized the leaves and roots of sugar beet plants following seed inoculation.