ISOLATION AND CHARACTERIZATION OF LINEAR DEOXYRIBONUCLEIC-ACID PLASMIDS FROM KLUYVEROMYCES-LACTIS AND THE PLASMID-ASSOCIATED KILLER CHARACTER
ISOLATION AND CHARACTERIZATION OF LINEAR DEOXYRIBONUCLEIC-ACID PLASMIDS FROM KLUYVEROMYCES-LACTIS AND THE PLASMID-ASSOCIATED KILLER CHARACTER
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DOI：
10.1128/jb.145.1.382-390.1981
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发表时间：
1981-01-01
影响因子：
3.2
通讯作者：
SAKAGUCHI, K
中科院分区：
文献类型：
--
作者：
GUNGE, N;TAMARU, A;SAKAGUCHI, K
从酵母K.lactis IFO 1267中分离到两个线形DNA质粒，命名为pGK11和pGKl2。PGK11和pGK12的相对分子质量为5.4倍。106和8.4倍。分别为106个。两种载体的密度均为1.687 g/cm~3，轻于线粒体(1.692 g/cm~3)和核DNA(1.699 g/cm~3)。用EcoRI、HindIII、PstI和BamHI酶切构建了pGKl1的限制性内切酶图谱。PGK12被EcoRI切割成7个片段，BamHI被切割成2个片段。乳酸克雷伯氏菌IFO 1267对酿酒酵母敏感和致命菌株以及某些菌株意大利葡萄球菌、乳酸克雷伯氏菌、耐热葡萄球菌和万古氏克雷伯氏菌均有杀灭作用。所有缺失pGK1质粒的乳酸克雷伯氏菌都是非杀菌剂。将乳酸克雷伯菌IFO 1267与一株非致死性乳酸克雷伯氏菌进行杂交，获得了产孢子后的四分体分析结果。杀手特征在所有四分体中与pGK1质粒相关联地以染色体外的方式传递。在任何一株乳酸克雷伯氏菌中均未检测到双链RNA杀伤子。杀人者的特征显然是由线状DNA质粒介导的。发现了一个单一的染色体基因，该基因对乳酸克雷伯氏菌具有抗药性。
Two linear DNA plasmids, designated pGKll and pGKl2, were isolated from the yeast K. lactis IFO 1267. pGKll and pGKl2 had MW of 5.4 .times. 106 and 8.4 .times. 106, respectively. Both plasmids possessed the same density of 1.687 g/cm3, lighter than the densities of mitochondrial (1.692 g/cm3) and nuclear (1.699 g/cm3) DNA. A restriction map of pGKl1 was constructed from digestions by EcoRI, HindIII, PstI, and BamHI. pGKl2 was cleaved by EcoRI into 7 fragments and by BamHI into 2 fragments. K. lactis IFO 1267 killed Saccharomyces cerevisiae sensitive and killer strains and certain strains of S. italicus, K. lactis, K. thermotolerans, and K. vanudenii. All K. lactis strains lacking the pGKl plasmids were nonkillers. A hybrid was constructed between K. lactis IFO 1267 and a nonkiller K. lactis strain lacking the plasmids and subjected to tetrad analysis after sporulation. The killer character was extrachromosomally transmitted in all tetrads in association with the pGKl plasmids. The double-stranded RNA killer plasmid could not be detected in any K. lactis killer strains. The killer character was apparently mediated by the linear DNA plasmids. A single chromosomal gene was found which was responsible for the resistance to the K. lactis killer.