Tinnell W H, Macrina F L
Infect Immun. 1976 Oct;14(4):955-64. doi: 10.1128/iai.14.4.955-964.1976.
Previous nucleic acid association studies have identified at least nine deoxyribonucleic acid (DNA) homology classes of the Bacteroides fragilis group of organisms. Using these classes as a taxonomic framework, we have screened representative strains of the B. fragilis group for the presence of extrachromosomal (plasmid) DNA. [3H]thymidine-labeled cell lysates were subjected to sodium dodecyl sulfate-salt precipitation, and supernatant fractions from such preparations were analyzed using cesium chloride-ethidium bromide equilibrium centrifugation. One strain from each group was examined in this fashion. Five of the strains were judged to contain no detectable plasmid DNA; however, four strains were observed to yield satellite bands corresponding to covalently closed circular plasmid DNA. Plasmid DNA from such gradients was subjected to velocity sedimentation through both neutral and alkaline sucrose gradients to determine molecular size. A 23 X 10(6)-molecular-weight plasmid was found in a B. fragilis strain representing one DNA homology group of this species, whereas a 3 X 10(6)-molecular-weight plasmid was found in a B. fragilis strain representing a second homology group. Similarly, a 31 X 10(6)-molecular-weight plasmid was found in a Bacteroides thetaiotaomicron strain representing one DNA homology group of this species, whereas a 3 X 10(6)-molecular-weight plasmid was found in a B. thetaiotaomicron strain representing a second homology group. In all instances, the small-molecular weight plasmids were present to the extent of about 15 copies per chromosomal equivalent, whereas the large plasmids were present to the extent of approximately 1 copy per chromosomal equivalent. The biological function of these plasmids is unknown.
先前的核酸相关性研究已经确定了脆弱拟杆菌属生物至少9个脱氧核糖核酸(DNA)同源类别。以这些类别作为分类框架,我们筛选了脆弱拟杆菌属的代表性菌株,以检测其是否存在染色体外(质粒)DNA。将[3H]胸苷标记的细胞裂解物进行十二烷基硫酸钠-盐沉淀,并用氯化铯-溴化乙锭平衡离心法分析此类制备物的上清液部分。每组选取一个菌株以此方式进行检测。其中5个菌株被判定不含可检测到的质粒DNA;然而,观察到4个菌株产生了与共价闭合环状质粒DNA相对应的卫星带。将来自此类梯度的质粒DNA通过中性和碱性蔗糖梯度进行速度沉降,以确定分子大小。在代表该物种一个DNA同源组的脆弱拟杆菌菌株中发现了一个分子量为23×10⁶的质粒,而在代表第二个同源组的脆弱拟杆菌菌株中发现了一个分子量为3×10⁶的质粒。同样,在代表该物种一个DNA同源组的多形拟杆菌菌株中发现了一个分子量为31×10⁶的质粒,而在代表第二个同源组的多形拟杆菌菌株中发现了一个分子量为3×10⁶的质粒。在所有情况下,小分子质量质粒的存在量约为每个染色体当量15份,而大质粒的存在量约为每个染色体当量1份。这些质粒的生物学功能尚不清楚。