Kato J, Amie J, Murata Y, Kuroda A, Mitsutani A, Ohtake H
Department of Fermentation Technology, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8527, Japan.
Appl Environ Microbiol. 1998 Jun;64(6):2061-4. doi: 10.1128/AEM.64.6.2061-2064.1998.
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 10(6) transformants per microg 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.
从沿海海水样本中分离出了四种能裂解硅藻中肋骨条藻NIES-324的海洋细菌，分别为交替单胞菌属菌株A27、A28、A29和A30。它们还能够裂解硅藻海链藻属和黄道营等鞭金藻，以及针胞藻属的古氏尖尾藻。在交替单胞菌属菌株A28和A29中分别检测到了隐蔽的内源质粒，命名为pAS28和pAS29。基于大小和限制性酶切位点分析，这些质粒似乎相似。通过将pAS28与大肠杆菌载体pCRIIc融合，构建了一种能在大肠杆菌和交替单胞菌属菌株A28中复制的穿梭载体。这个16千碱基对的嵌合质粒命名为pASS1，其转化菌株A28的频率为每微克DNA产生10⁶个转化子。对pASS1的缺失分析表明，pAS28的4.7千碱基对EcoRI-HindIII区域对于在菌株A28中维持质粒至关重要。这个EcoRI-HindIII片段包含一个开放阅读框，似乎编码一种708个氨基酸的蛋白质。
