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Development of a Genetic Transformation System for an Alga-Lysing Bacterium

机译:溶藻细菌遗传转化系统的开发

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摘要

Four marine bacteria, Alteromonas sp. strains A27, A28, A29, and A30, that lyse the diatom Skeletonema costatum NIES-324 were isolated from coastal seawater samples. They were also able to lyse the diatoms Thalassiosira sp. and Eucampia zodiacs and the raphidophycean flagellate Chattonella antiqua. Cryptic indigenous plasmids, designated pAS28 and pAS29, were detected in Alteromonas sp. strains A28 and A29, respectively. These plasmids appeared to be similar based on size and restriction site analysis. A shuttle vector that replicates in Escherichia coli and Alteromonas sp. strain A28 was constructed by fusing pAS28 and E. coli vector pCRIIc. The 16-kbp chimeric plasmid, designated pASS1, had the ability to transform strain A28 at a frequency of 106 transformants per μg of DNA. Deletion analysis of pASS1 showed that the 4.7-kb EcoRI-HindIII region of pAS28 was essential for plasmid maintenance in strain A28. This EcoRI-HindIII fragment contained an open reading frame which appeared to encode a 708-amino-acid protein.
机译:四种海洋细菌,Alteromonas sp。从沿海海水样本中分离了裂解硅藻骨架骨骼NIES-324的A27,A28,A29和A30菌株。他们还能够裂解硅藻Thalassiosira sp。和黄柏黄道虫和鳞茎藻鞭毛产的产查通氏菌。在Alteromonas sp。中检测到了隐性原生质粒,命名为pAS28和pAS29。分别为A28和A29菌株。基于大小和限制性位点分析,这些质粒看起来相似。在大肠杆菌和Alteromonas sp。中复制的穿梭载体。通过融合pAS28和大肠杆菌载体pCRIIc来构建菌株A28。 16 kbp嵌合质粒称为pASS1,具有以每μgDNA 10 6 个转化子的频率转化A28菌株的能力。 pASS1的缺失分析表明,pAS28的4.7-kb EcoRI-HindIII区对于菌株A28中的质粒维持至关重要。此EcoRI-HindIII片段含有一个开放阅读框,似乎编码一个708个氨基酸的蛋白质。

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