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通过重复外显子回文序列-聚合酶链反应、16S rDNA测序和傅里叶变换红外光谱表征,确定用金氏B培养基和古尔德S1琼脂分离的假单胞菌菌株的多样性。

Diversity of Pseudomonas strains isolated with King's B and Gould's S1 agar determined by repetitive extragenic palindromic-polymerase chain reaction, 16S rDNA sequencing and Fourier transform infrared spectroscopy characterisation.

作者信息

Johnsen K, Nielsen P

机构信息

Department of Ecology, Royal Veterinary and Agricultural University, Copenhagen, Denmark.

出版信息

FEMS Microbiol Lett. 1999 Apr 1;173(1):155-62. doi: 10.1111/j.1574-6968.1999.tb13497.x.

Abstract

King's B and Gould's S1 agar were compared with regard to the isolation of Pseudomonas from four environmental samples. In all samples, King's B gave the highest number of colony-forming units, and in some environments, there were more fluorescent colony-forming units on King's B as well. However, almost all types grew on Gould's S1, which enabled us to isolate a greater variety of groups than with King's B, fluorescent as well as non-fluorescent members of Pseudomonas. The Pseudomonas isolates were comparatively typed by repetitive extragenic palindromic-polymerase chain reaction and Fourier transform infrared spectroscopy, not previously used for environmental Pseudomonas. The two typing methods were similar in resolution, thus Fourier transform infrared spectroscopy proved fast and reproducible and is a good method for discrimination at subspecies level. Representative strains were identified by partial 16S rDNA sequencing. Thus, we suggest Gould's S1 agar be used for isolation of Pseudomonas because the results are reproducible, specific and give the most diverse recovery and the least work.

摘要

就从四个环境样本中分离假单胞菌而言,对金氏B培养基和古尔德S1琼脂进行了比较。在所有样本中,金氏B培养基产生的菌落形成单位数量最多,而且在某些环境中,金氏B培养基上的荧光菌落形成单位也更多。然而,几乎所有类型的假单胞菌都能在古尔德S1琼脂上生长,这使我们能够分离出比使用金氏B培养基更多种类的菌群,包括荧光和非荧光的假单胞菌成员。通过重复外源性回文序列聚合酶链反应和傅里叶变换红外光谱法对分离出的假单胞菌进行分型,这两种方法以前未用于环境中的假单胞菌。这两种分型方法在分辨率上相似,因此傅里叶变换红外光谱法证明快速且可重复,是在亚种水平上进行区分的好方法。通过部分16S rDNA测序对代表性菌株进行鉴定。因此,我们建议使用古尔德S1琼脂来分离假单胞菌,因为其结果可重复、特异,能实现最多样化的回收且工作量最小。

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