scholarly journals PCR amplification of species specific sequences of 16S rDNA and 16S–23S rDNA intergenic spacer region for identification of Streptococcus phocae

2008 ◽  
Vol 163 (2) ◽  
pp. 132-135 ◽  
Author(s):  
A.A. Hassan ◽  
A. Vossen ◽  
C. Lämmler ◽  
U. Siebert ◽  
J.F. Fernández-Garayzábal
Keyword(s):  
16S Rdna ◽  
Pcr Amplification ◽  
Intergenic Spacer ◽  
Intergenic Spacer Region ◽  
Rdna Intergenic Spacer ◽  
Species Specific ◽  
Specific Sequences

Sequence Length Variation of Internal Genic Space of 16S rDNA-23S rDNA in Bacterium with High Yield of Hydrogen Production

2011 ◽  
Vol 183-185 ◽  
pp. 1413-1416
Author(s):  
Yong Feng Li ◽  
Yi Xuan Wang ◽  
Lu Wang ◽  
Zhan Qing Wang
Keyword(s):  
16S Rdna ◽  
Pcr Amplification ◽  
Intergenic Spacer ◽  
Rrna Genes ◽  
High Yield ◽  
23S Rrna ◽  
Sequence Length ◽  
Rrna Gene ◽  
Length Variation ◽  
Intergenic Spacer Region

To develop the identification of species for fermentative biohydrogen-producing bacterium, scholars have found a method which is based on PCR amplification of the 16S rRNA gene (rDNA)-23S rDNA intergenic regions. In the study, a large fragment of the rDNA operon, including the 16S rDNA, the intergenic spacer region (ISR) and approximately 2000 bases of the 23S rDNA, were polymerasechain reaction (PCR) amplified. The PCR amplification of the genomic DNA of Leptonema ilk strain 3055 using primers directed against conserved regions of the rRNA operon provided evidence that the 16S and 23S rRNA genes were linked via an intergenic spacer region. The sequencing of the intergenic spacer region indicated that it was 435 nucleotides in length and sequence similarity searches revealed that it bore no homology to any known sequences including tRNA available in databases.

scholarly journals Identification of Nile Perch (Lates niloticus), Grouper (Epinephelus guaza), and Wreck Fish (Polyprion americanus) Fillets by PCR Amplification of the 5S rDNA Gene

2001 ◽  
Vol 84 (3) ◽  
pp. 777-781 ◽  
Author(s):  
Luis Asensio ◽  
Isabel González ◽  
Alicia Fernández ◽  
Ana Céspedes ◽  
Miguel A Rodríguez ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Intergenic Spacer ◽  
5S Rdna ◽  
Nile Perch ◽  
Intergenic Spacer Region ◽  
Rdna Gene ◽  
Lates Niloticus ◽  
Polymerase Chain ◽  
Species Specific ◽  
Polyprion Americanus

Abstract Nile perch (Lates niloticus), grouper (Epinephelus guaza), and wreck fish (Polyprion americanus) were differentiated by polymerase chain reaction (PCR) amplification of the nuclear 5S rDNA gene. The design of 3 species-specific primers complementary to the nontranscribed intergenic spacer region from the 5S rDNA molecule allowed amplification of clearly distinguishable gene fragments in each fish species. This approach is rapid and reliable and offers the potential to detect fraudulent or unintentional mislabeling of these species in routine seafood authentication analysis.

scholarly journals PCR amplification and polymorphism analysis of the intergenic spacer region of ribosomal DNA in Tuber borchii

2002 ◽  
Vol 157 (1) ◽  
pp. 69-74 ◽  
Author(s):  
Pasquapina Ciarmela ◽  
Lucia Potenza ◽  
Luigi Cucchiarini ◽  
Sabrina Zeppa ◽  
Vilberto Stocchi
Keyword(s):  
Ribosomal Dna ◽  
Pcr Amplification ◽  
Intergenic Spacer ◽  
Polymorphism Analysis ◽  
Tuber Borchii ◽  
Intergenic Spacer Region

scholarly journals Evaluation of Nonpathogenic Fusarium oxysporum and Pseudomonas fluorescens for Panama Disease Control

Plant Disease ◽  
2011 ◽  
Vol 95 (8) ◽  
pp. 951-959 ◽  
Author(s):  
A. Belgrove ◽  
C. Steinberg ◽  
A. Viljoen
Keyword(s):  
Fusarium Oxysporum ◽  
Pseudomonas Fluorescens ◽  
Fusarium Wilt ◽  
Intergenic Spacer ◽  
Polymorphism Analysis ◽  
Panama Disease ◽  
Intergenic Spacer Region ◽  
Soil Microbial ◽  
Fusarium Wilt Of Banana ◽  
Species Specific

Nonpathogenic Fusarium oxysporum endophytes from healthy banana roots were evaluated for their ability to reduce Fusarium wilt of banana (Panama disease). Isolates were identified morphologically and by using species-specific primers. Pathogenicity was confirmed by inoculating banana plantlets in the greenhouse. Nonpathogenic F. oxysporum isolates were grouped into 14 haplotype groups by polymerase chain reaction restriction fragment length polymorphism analysis of the intergenic spacer region, and representative isolates evaluated for biocontrol of F. oxysporum f. sp. cubense. In the greenhouse, 10 nonpathogenic F. oxysporum isolates were able to significantly reduce Fusarium wilt of banana. The isolate that protected banana plantlets best in the greenhouse, a nonpathogenic F. oxysporum from the root rhizosphere, and Pseudomonas fluorescens WCS 417 were then field tested. When the putative biological control organisms were tested in the field, neither the nonpathogenic F. oxysporum, P. fluorescens, nor combinations thereof reduced Fusarium wilt development significantly. A number of factors could contribute to the lack of field protection, including soil microbial and chemical composition and reduced survival of biocontrol organisms in banana roots. A lack of knowledge regarding the etiology of Fusarium wilt of ‘Cavendish’ banana in the subtropics and the effect of F. oxysporum f. sp. cubense race and banana cultivar in protection of banana by biocontrol organisms should be further investigated.

scholarly journals Identification of Porphyromonas gingivalis Strains by Heteroduplex Analysis and Detection of Multiple Strains

1999 ◽  
Vol 37 (12) ◽  
pp. 3906-3911 ◽  
Author(s):  
Eugene J. Leys ◽  
James H. Smith ◽  
Sharon R. Lyons ◽  
Ann L. Griffen
Keyword(s):  
Porphyromonas Gingivalis ◽  
Dna Sequences ◽  
Allelic Variation ◽  
Intergenic Spacer ◽  
Heteroduplex Analysis ◽  
Clinical Samples ◽  
Specific Primers ◽  
Intergenic Spacer Region ◽  
Species Specific ◽  
Multiple Strains

Heteroduplex analysis has been used extensively to identify allelic variation among mammalian genes. It provides a rapid and reliable method for determining and cataloging minor differences between two closely related DNA sequences. We have adapted this technique to distinguish among strains or clonal types of Porphyromonas gingivalis. The ribosomal intergenic spacer region (ISR) was amplified directly from a subgingival plaque sample by PCR with species-specific primers, avoiding the need for culturing the bacteria. The PCR products were then directly compared by heteroduplex analysis with known strains of P. gingivalis for identification. We identified 22 distinct but closely related heteroduplex types ofP. gingivalis in 1,183 clinical samples. Multiple strains were found in 34% of the samples in which P. gingivaliswas detected. Heteroduplex types were identified from these multistrain samples without separating them by culturing or molecular cloning. PCR with species-specific primers and heteroduplex analysis makes it possible to reliably and sensitively detect and identify strains ofP. gingivalis in large numbers of samples.

PCR Detection Assay of Fumonisin-Producing Fusarium verticillioides Strains

2004 ◽  
Vol 67 (6) ◽  
pp. 1278-1283 ◽  
Author(s):  
BELÉN PATIÑO ◽  
SALVADOR MIRETE ◽  
M. TERESA GONZÁLEZ-JAÉN ◽  
GIUSEPPINA MULÉ ◽  
M. TERESA RODRÍGUEZ ◽  
...  
Keyword(s):  
Fusarium Species ◽  
Fusarium Verticillioides ◽  
Animal Health ◽  
Intergenic Spacer ◽  
Fungal Species ◽  
Intergenic Spacer Region ◽  
Detection And Identification ◽  
Geographical Regions ◽  
Species Specific ◽  
A Minor

Fusarium verticillioides is considered to be the main source of fumonisins, a group of toxins that contaminate commodities and result in chronic and acute diseases affecting humans and animals. The detection and control of this species is crucial to prevent fumonisins from entering the food chain. The objective of the present research was to develop a specific, sensitive, and robust PCR assay to detect F. verticillioides strains using two pairs of specific primers for F. verticillioides, which have been designed on the basis of the intergenic spacer region of the rDNA units. The first pair of primers was F. verticillioides species specific, whereas the second pair of primers detected fumonisin-producing F. verticillioides strains. This second pair of primers allowed for the discrimination between the major group of F. verticillioides strains, fumonisin-producing strains that are mainly associated with crops, and a minor group of strains, non–fumonisin-producing strains that are associated with bananas. Fifty-four strains of F. verticillioides from different geographical regions and hosts were tested using both sets of primers. Sixteen additional Fusarium species were examined. The specificity of the primer sequences provides the basis for a simple, rapid, accurate, and sensitive detection and identification method of this fungal species that represents a risk for human and animal health.

scholarly journals Species-Specific Plasmid Sequences for PCR Identification of the Three Species of Borrelia burgdorferiSensu Lato Involved in Lyme Disease

1998 ◽  
Vol 36 (1) ◽  
pp. 269-272 ◽  
Author(s):  
Marie-Christine Misonne ◽  
Philippe Pierre Hoet
Keyword(s):  
Lyme Disease ◽  
Pcr Amplification ◽  
Sensu Stricto ◽  
Pcr Assay ◽  
Borrelia Garinii ◽  
Specific Pcr ◽  
Pcr Identification ◽  
Identification Of Species ◽  
Species Specific ◽  
Specific Sequences

Species-specific sequences were shown to be carried by plasmids of the three main species of Borrelia burgdorferi sensu lato involved in Lyme disease. Libraries of the 16-, 33-, and 25-kb plasmids of B. burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii, respectively, were then built and used to isolate species-specific sequences. After sequencing of the cloned inserts, three sets of primers were designed. They were shown to determine species-specific PCR amplification products. The sensitivities of the PCR assay with these primers were 100 spirochetes for B. burgdorferi sensu stricto and 1,000 spirochetes for B. garinii and B. afzelii. The usefulness of these primers for the identification of species in biological samples (tick, serum, and cerebrospinal fluid samples) was ascertained.

Sign in / Sign up

Export Citation Format

Share Document