Zhu H, Zhou W Y, Xu M, Shen Y L, Wei D Z
State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, P.R. China.
Lett Appl Microbiol. 2007 Aug;45(2):174-8. doi: 10.1111/j.1472-765X.2007.02166.x.
To establish the specific DNA patterns in 16S rDNA and 16S-23S rDNA intergenic spacer (IGS) regions from different kinds of Serratia marcescens strains using polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP) and sequences analysis.
Two pairs of primers based on the 16S rDNA and 16S-23S rDNA IGS were applied to amplify the rrn operons of two kinds of S. marcescens strains. About 1500 bp for 16S rDNA and four fragments of different sizes for 16S-23S rDNA IGS were obtained. PCR-amplified fragments were analysed by RFLP and sequence analysis. Two distinct restriction patterns revealing three to five bands between two kinds of strains were detected with each specific enzyme. According to the sequence analysis, two kinds of strains showed approximately 97% sequence homology of 16S rDNA. However, there was much difference in the sequences of IGS between the two kinds of strains. Intercistronic tRNA of strains H3010 and A3 demonstrated an order of tRNA of 5'-16S-tRNA(Ala)-tRNA(Ile)-23S-3', but strain B17 harboured the tRNA of 5'-16S-tRNA(Glu)-tRNA(Ile)-23S-3'.
The method was specific, sensitive and accurate, providing a new technique for differentiating different strains from the same species.
This paper provided the first molecular characterization of 16S rDNA and 16S-23S rDNA IGS from S. marcescens strains.
运用聚合酶链反应（PCR）、限制性片段长度多态性（RFLP）及序列分析，确定不同种类粘质沙雷氏菌菌株16S核糖体DNA（rDNA）和16S - 23S rDNA基因间隔区（IGS）的特定DNA模式。
应用基于16S rDNA和16S - 23S rDNA IGS的两对引物，扩增两种粘质沙雷氏菌菌株的rrn操纵子。获得了约1500 bp的16S rDNA以及16S - 23S rDNA IGS的四个不同大小的片段。对PCR扩增片段进行RFLP和序列分析。用每种特异性酶检测到两种菌株之间有两种不同的限制性模式，显示出三到五条带。根据序列分析，两种菌株的16S rDNA序列同源性约为97%。然而，两种菌株IGS的序列存在很大差异。菌株H3010和A3的顺反子间tRNA呈现5'-16S-tRNA（丙氨酸）-tRNA（异亮氨酸）-23S-3'的tRNA顺序，但菌株B17含有5'-16S-tRNA（谷氨酸）-tRNA（异亮氨酸）-23S-3'的tRNA。
该方法特异、灵敏且准确，为区分同一物种的不同菌株提供了一种新技术。
本文首次对粘质沙雷氏菌菌株的16S rDNA和16S - 23S rDNA IGS进行了分子特征分析。
