Quinlan D C, Maniloff J
J Bacteriol. 1972 Dec;112(3):1375-9. doi: 10.1128/jb.112.3.1375-1379.1972.
Newly replicated deoxyribonucleic acid (DNA) in Mycoplasma gallisepticum A5969 is membrane associated. Cells pulse-labeled 1 to 3 min with (3)H-thymidine are lysed by a freeze-thaw procedure. After brief sonic treatment to shear the DNA, differential centrifugation gives a cell fraction (P2) that is enriched sevenfold for pulse-labeled DNA. P2 contains 80% of the total adenosine triphosphatase activity, 65% of the total cholesterol, and morphologically intact terminal bleb structures. Three to four minutes are needed to fully label the DNA growing-point region, whereas 7 to 8 min are required to "chase" 50% of the (3)H-labeled DNA. This pulse-chase removes 80 to 85% of the nascent DNA from the P2 fraction.
鸡毒支原体A5969中新复制的脱氧核糖核酸(DNA)与细胞膜相关。用(3)H-胸腺嘧啶脉冲标记1至3分钟的细胞通过冻融程序裂解。经过短暂的超声处理以剪切DNA后,差速离心得到一个细胞组分(P2),其脉冲标记的DNA富集了七倍。P2含有总腺苷三磷酸酶活性的80%,总胆固醇的65%,以及形态完整的末端泡状结构。完全标记DNA生长点区域需要3至4分钟,而“追踪”50%的(3)H标记的DNA则需要7至8分钟。这种脉冲追踪从P2组分中去除了80%至85%的新生DNA。