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通过差示扫描量热法测定的细菌DNA碱基组成与其细胞内热解链温度之间的关系。

The relationship between the base composition of bacterial DNA and its intracellular melting temperature as determined by differential scanning calorimetry.

作者信息

Mackey B M, Parsons S E, Miles C A, Owen R J

机构信息

AFRC Institute of Food Research, Bristol Laboratory, Langford, UK.

出版信息

J Gen Microbiol. 1988 May;134(5):1185-95. doi: 10.1099/00221287-134-5-1185.

Abstract

The correlation between the melting temperature of intracellular DNA, determined by differential scanning calorimetry (DSC) of whole bacteria, and its guanine + cytosine (G + C) content, was examined for 58 species of bacteria. Samples of vegetative cells were heated in a Perkin-Elmer DSC-2C at 10 degrees C min-1 from 5 to 130 degrees C, cooled to 5 degrees C and then re-heated as before. Literature values for the mole fraction of G + C, XGC, were linearly related to the temperature, Tmax, at which the reversible peak, pr, observed on the second heating run was at a maximum, via the equation XGC = (Tmax -73.8)/41.0. This equation accounted for 91.9% of the variance in XGC with 95% confidence limits of +/- 7.3%, approximately 1.6 times the corresponding uncertainty (+/- 4.5%) quoted by De Ley (Journal of Bacteriology 101, 738-754, 1970) for estimates based on the spectroscopically determined melting temperature of purified DNA. Random errors of measurement of Tmax did not greatly limit the precision of the prediction and it was concluded that factors additional to base composition affected the temperature of DNA melting within the bacterial cell. Displacement of Tmax values from the fitted line was particularly noticeable in Campylobacter, Corynebacterium and Bacterionema species and part of the residual variation appeared to be species specific, possibly caused by differences in intracellular solute concentration.

摘要

通过对58种细菌进行全菌差示扫描量热法(DSC)测定,研究了细胞内DNA的解链温度与其鸟嘌呤+胞嘧啶(G + C)含量之间的相关性。将营养细胞样品在珀金埃尔默DSC - 2C中以10℃/分钟的速率从5℃加热至130℃,冷却至5℃,然后如前所述再次加热。G + C摩尔分数XGC的文献值与第二次加热运行时观察到的可逆峰pr最大值时的温度Tmax呈线性关系,其方程为XGC = (Tmax - 73.8)/41.0。该方程解释了XGC中91.9%的方差,95%置信限为±7.3%,约为德莱(《细菌学杂志》101, 738 - 754, 1970)基于光谱测定的纯化DNA解链温度所引用的相应不确定性(±4.5%)的1.6倍。Tmax测量的随机误差并未严重限制预测的精度,得出的结论是,除碱基组成外的其他因素会影响细菌细胞内DNA的解链温度。在弯曲杆菌属、棒状杆菌属和纤毛菌属物种中,Tmax值偏离拟合线的情况尤为明显,部分残余变异似乎具有物种特异性,可能是由细胞内溶质浓度差异引起的。

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