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Gyrase activity and number of copies of the gyrase B subunit gene in Haemophilus influenzae.

机译:流感嗜血杆菌中的回旋酶活性和回旋酶B亚基基因的拷贝数。

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摘要

Gyrase activities in extracts of various strains of Haemophilus influenzae can differ by more than an order of magnitude (J. K. Setlow, E. Cabrera-Juárez, W. L. Albritton, D. Spikes, and A. Mutschler, J. Bacteriol. 164:525-534, 1985). Measurements of in vitro activity and copy number indicated that most of these differences arose from variations in the number of copies of the gene for the gyrase B subunit, with some strains containing multicopy plasmids coding for that subunit. The quantitative relationship between gyrase and copy number depended on the mutations in the plasmids and in the host. The gyrase and copy number were considerably lower in plasmid-bearing strains carrying the prophage HP1c1. Two mutations affecting gyrase that are apparently regulatory caused an increase in gyrase without a concomitant increase in copy number. The possibility that the in vivo gyrase activity did not reflect the in vitro data was explored by measurement of alkaline phosphatase and ATPase activity in the extracts. Alkaline phosphatase activity increased with increasing gyrase activity measured in vitro, but ATPase activity did not. We conclude that extra supercoiling enhanced transcription of the alkaline phosphatase gene but not the ATPase gene and that it is unlikely that there is much discrepancy between gyrase activity assayed in vitro and the activity in the cell.
机译:各种流感嗜血杆菌菌株提取物中的促旋酶活性可能相差一个数量级以上(JK Setlow,E.Cabrera-Juárez,WL Albritton,D.Spikes,and A.Mutschler,J.Bacteriol。164:525-534 (1985年)。体外活性和拷贝数的测量结果表明,这些差异中的大多数是由回旋酶B亚基的基因拷贝数变化引起的,某些菌株含有编码该亚基的多拷贝质粒。促旋酶和拷贝数之间的定量关系取决于质粒和宿主中的突变。在携带原噬菌体HP1c1的带有质粒的菌株中,回旋酶和拷贝数要低得多。明显影响调节的两个影响旋转酶的突变引起了旋转酶的增加,而拷贝数却没有随之增加。通过测量提取物中碱性磷酸酶和ATP酶的活性,探索了体内促旋酶活性不能反映体外数据的可能性。碱性磷酸酶活性随体外测量的回旋酶活性的增加而增加,但ATPase活性却没有。我们得出的结论是,额外的超螺旋增强了碱性磷酸酶基因的转录,但没有增强ATPase基因的转录,并且在体外测定的回旋酶活性与细胞活性之间不可能存在很大差异。

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