Evaluation of the rrn operon copy number in Bifidobacterium using real-time PCR

被引：29

作者：

Candela, M ^{[1
]}

Vitali, B ^{[1
]}

Matteuzzi, D ^{[1
]}

Brigidi, P ^{[1
]}

机构：

[1] Univ Bologna, Dept Pharmaceut Sci, CIRB Ctr Biotechnol, I-40126 Bologna, Italy

来源：

LETTERS IN APPLIED MICROBIOLOGY | 2004年 / 38卷 / 03期

关键词：

Bifidobacterium; gene quantification; probiotics; real-time PCR; ribosomal operon redundancy;

D O I：

10.1111/j.1472-765X.2003.01475.x

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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.

引用

页码：229 / 232

页数：4

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