Evaluation of the rrn operon copy number in Bifidobacterium using real-time PCR
Evaluation of the rrn operon copy number in Bifidobacterium using real-time PCR
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DOI：
10.1111/j.1472-765x.2003.01475.x
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发表时间：
2004-01-01
影响因子：
2.4
通讯作者：
Brigidi, P
中科院分区：
文献类型：
--
作者：
Candela, M;Vitali, B;Brigidi, P
目的：建立一种基于实时荧光PCR的双歧杆菌rRNA操纵子拷贝数检测方法。由于其重复的性质，rRNA操纵子是非常合适的目标，染色体整合的异源genes.Methods和结果：rrn操纵子的多重性每个染色体的16S rRNA扩增子从基因组DNA获得的实时PCR定量测定。在几种人源双歧杆菌菌株中获得的值范围为1至5。Southern杂交技术证实了该方法的可靠性。结论：在双歧杆菌属中，rrn操纵子拷贝数在种和菌株水平上存在变异性。双歧杆菌菌株具有高rRNA多样性的鉴定允许选择潜在的主机染色体integration.Significance和影响的研究：这里提出的方法是一个快速，可靠和灵敏的新工具rrn操纵子拷贝数在细菌中的定量。
Aims: A real-time PCR-based method was developed to evaluate the Bifidobacterium rRNA operon copy number. As a result of their repetitive nature, rRNA operons are very suitable targets for chromosomal integration of heterologous genes.Methods and Results: The rrn operon multiplicity per chromosome was determined by real-time PCR quantification of the 16S rRNA amplicons obtained from genomic DNA. The values obtained in several bifidobacterial strains of human origin ranged from 1 to 5. The reliability of the method developed was confirmed by Southern hybridization technique.Conclusions: In the Bifidobacterium genus the rrn operon copies showed variability at species and strain level. The identification of Bifidobacterium strains with high rRNA multiplicity allowed the selection of potential hosts for chromosomal integration.Significance and Impact of the Study: The methodology here proposed represents a rapid, reliable and sensitive new tool for the quantification of rrn operon copy number in bacteria.