Salyers A A, Guthrie E P
Department of Microbiology, University of Illinois, Urbana 61801.
Appl Environ Microbiol. 1988 Aug;54(8):1964-9. doi: 10.1128/aem.54.8.1964-1969.1988.
Bacteroides thetaiotaomicron, an obligate anaerobe normally found in high concentrations in the human colon, is one of the few colon bacteria that can ferment host mucopolysaccharides such as chondroitin sulfate. Previously, we found that a directed insertional mutation in the gene that codes for the chondroitinase II gene of B. thetaiotaomicron did not affect growth on chondroitin sulfate despite the fact that chondroitinase II accounts for 70% of the total cellular chondroitinase activity. Thus, the chondroitinase II gene did not seem to contribute significantly to growth on chondroitin sulfate when the bacteria were grown in laboratory medium. To determine whether this enzyme is important for bacteria growing in the intestinal tract, we tested the ability of a strain that does not produce chondroitinase II to colonize the intestinal tracts of germfree mice and to compete with wild-type B. thetaiotaomicron. The mutant used in these experiments carried a 0.5-kilobase deletion in the chondroitinase II gene and was constructed so that, unlike the original insertion mutant, it contained no exogenous DNA. The deletion mutant colonized the intestinal tracts of germfree mice at the same levels as the wild type. When a mixture of the deletion mutant and wild type was used to colonize germfree mice, the percent wild type, measured by colony hybridization with the deleted 0.5-kilobase fragment as the hybridization probe, did not rise to 100% even after periods as long as 9 weeks. In most experiments, the percent wild type did not rise significantly above the percent in the original mixture.(ABSTRACT TRUNCATED AT 250 WORDS)
多形拟杆菌是一种专性厌氧菌,通常在人类结肠中大量存在,是少数能够发酵宿主黏多糖(如硫酸软骨素)的结肠细菌之一。此前,我们发现,多形拟杆菌中编码硫酸软骨素酶II基因的定向插入突变,尽管硫酸软骨素酶II占细胞总硫酸软骨素酶活性的70%,但并不影响其在硫酸软骨素上的生长。因此,当细菌在实验室培养基中生长时,硫酸软骨素酶II基因似乎对在硫酸软骨素上的生长没有显著贡献。为了确定这种酶对在肠道中生长的细菌是否重要,我们测试了一株不产生硫酸软骨素酶II的菌株在无菌小鼠肠道中定殖以及与野生型多形拟杆菌竞争的能力。这些实验中使用的突变体在硫酸软骨素酶II基因上有一个0.5千碱基的缺失,并且构建方式使得与原始插入突变体不同,它不包含外源DNA。缺失突变体在无菌小鼠肠道中的定殖水平与野生型相同。当使用缺失突变体和野生型的混合物对无菌小鼠进行定殖时,以缺失的0.5千碱基片段作为杂交探针通过菌落杂交测量的野生型百分比,即使在长达9周的时间后也没有升至100%。在大多数实验中,野生型百分比没有显著高于原始混合物中的百分比。(摘要截短于250字)