荧光法体内测定枯草芽孢杆菌和噬菌体2C的DNA。

Fluorometric in vivo determination of Bacillus subtilis and phage 2C DNA.

作者信息

Daxhelet G A, Kohnen M M, Coene M M, Hoet P P

机构信息

Microbiology and Genetics Unit, University of Louvain Medical School, Brussels, Belgium.

出版信息

Anal Biochem. 1990 Oct;190(1):116-9. doi: 10.1016/0003-2697(90)90143-w.

DOI:10.1016/0003-2697(90)90143-w

PMID:2126677

Abstract

The validity of in vivo fluorometric assays was ascertained for phage and bacterial DNA measurements. The following parameters were determined by this simple technique. The DNA content of dividing cells of Bacillus subtilis 168/2 was 2.65 times higher than in resting cells. Assuming that resting cells harbor 1 genomic equivalent, its Mr was estimated to be 4.4 x 10(9) Da. A polymerization rate during growth of 788,000 bp min-1/cell is accounted for by a multifork replication mechanism. Both phage and host DNA could be measured accurately during the lytic cycle. Phage 2C DNA synthesis proceeded at a linear rate of 5.2 genome equivalents min-1.

摘要

确定了体内荧光测定法在噬菌体和细菌DNA测量中的有效性。通过这种简单技术测定了以下参数。枯草芽孢杆菌168/2分裂细胞的DNA含量比静止细胞高2.65倍。假设静止细胞含有1个基因组当量，其相对分子质量估计为4.4×10⁹ Da。多叉复制机制解释了生长过程中788,000 bp min⁻¹/细胞的聚合速率。在裂解周期中，可以准确测量噬菌体和宿主DNA。噬菌体2C DNA合成以5.2个基因组当量min⁻¹的线性速率进行。