【摘要】 从国内多个地区的65份土样中分离纯化了产荧光假单胞菌株146株,其中菌株DSC3和DKXC1含有质粒。细胞形态、菌落特征的观察和检测证明DKXC1菌株为恶臭假单胞菌（Pseudomonas putida）。以其为出发菌株,克隆了该菌质粒DNA复制子1.7kb片段,序列分析结果表明该DNA片段由3个ORF组成,分别编码311、130、115个氨基酸;它与已知的Pf-E.coli穿梭质粒pUCP19的复制区同源性只有49％。该复制子DNA序列已在GenBank登记,登记号为AF273219,这是国内分离克隆的第一个Pseudomonas质粒DNA复制子。以此复制子构建了4.7kb的E.coli-Pf穿梭载体pYQSPE,该质粒在荧光假单胞菌P303中能够稳定复制和遗传。更多还原

【Abstract】 46 isolates of fluorescent Pseudomonas were obtained from different soil samples collected in several provinces. The strain, DKXC1, harboring 3.0 kb plasmid (pDX39), was identified as Pseudomonas putida. 1.7 kb plasmid DNA replicon of pDX39 was cloned and sequenced subsequently, three ORFs was found on the DNA and homology analysis showed that this replicon had low homology compared with the known replicon of pUCP19. It was inverted into pBluescriptSK+, and one new E.coli-Pseudomonas shuttle vector pYQSPE was obtained. pYQSPE was transformed into wild Pseudomonas fluorescence strain P303, and it was stable and could replicate normally. This replicon sequence had been registered in GenBank, and the accession number was AF273219.更多还原