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OALib Journal期刊
ISSN: 2333-9721
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费氏中华根瘤菌hn01的puta基因克隆
DOI: 10.3724/SP.J.1145.2010.00697, PP. 697-700
Keywords: 费氏中华根瘤菌hn01,puta基因,克隆,突变体,植株实验,结瘤
Abstract:
从快生型大豆根瘤菌费氏中华根瘤菌hn01的基因组文库中克隆到一个puta（脯氨酸脱氢酶）基因，该基因与已报道的苜蓿中华根瘤菌1021中puta基因在核苷酸和氨基酸水平上分别有82%和86%相似性.利用自杀性质粒pk18mob构建含有puta基因部分片段的重组质粒，通过三亲本结合导入出发菌株hn01中，获得正向插入的极性突变体gxhnpa和反向插入非极性突变体gxhnpb.将puta基因完整的orf连接到广宿主载体plafrj上，获得用于互补的质粒pgxhn37，通过三亲接合，将plafrj导入gxhnpa和gxhnpb获得互补菌株gxhnpha和gxhnphb.对出发菌株、突变菌株、互补菌株进一步的研究发现，以脯氨酸为唯一c、n源的mm培养基中培养时，突变体均不能生长，互补菌株和野生型hn01没有差异，而在完全培养基ymb中培养时，突变菌株生长不受影响.植株实验发现，突变体均能有效结瘤，但是固氮酶活与出发菌株相比有所下降，结瘤时间延迟1d，竞争结瘤能力下降，而互补菌株与野生型hn01没有明显的差异.图6表1参19
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