Restriction Enzyme Analysis of Campylobacter Genomic DNA

Streptomyces rochei A2 endoglucanase (eglS) and β-glucosidase (bgs1) genes were used as probes to survey their distribution among 16 Streptomyces strains isolated from different sources and characterized for their cellulolytic activities. The eglS probe hybridized to the genomic DNA of 12 strains with a restriction pattern different from that of S. rochei A2. The DNA from all strains, except one, hybridized with the bgsl probe and one strain showed the same restriction pattern as seen in S. rochei A2. The sequence localized by the eglS probe in S. thermoviolaceus and the one localized by the bgs1 probe in strain EC1 were cloned and expressed in E. coli in plasmids pTAE and pCSF203, respectively. The restriction maps showed that the cloned genes were identical to eglS and bgs1. The restriction enzyme analysis of genomic DNA from all the strains identified nine different groups, each characterized by a distinctive pattern and in agreement with the results of the hybridization experiments.Key words: Streptomyces, cellulase genes, hybridization, restriction enzyme analysis.

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Homozygosity for the transthyretin-Met30-gene in seven individuals with familial amyloidosis with polyneuropathy detected by restriction enzyme analysis of amplified genomic DNA sequences

Clinical Genetics ◽

10.1111/j.1399-0004.1992.tb03627.x ◽

2008 ◽

Vol 41 (1) ◽

pp. 39-41 ◽

Cited By ~ 36

Author(s):

Gōsta Holmgren ◽

Sven Bergström ◽

Ulf Drugge ◽

Erik Lundgren ◽

Carin Nording-Sikström ◽

...

Keyword(s):

Restriction Enzyme ◽

Dna Sequences ◽

Genomic Dna ◽

Enzyme Analysis ◽

Restriction Enzyme Analysis

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PCR-based Single-strand Conformation Polymorphism (SSCP) Analysis to Clone Nine Aquaporin Genes in Cucumber

Journal of the American Society for Horticultural Science ◽

10.21273/jashs.127.6.925 ◽

2002 ◽

Vol 127 (6) ◽

pp. 925-930 ◽

Cited By ~ 5

Author(s):

Jiahua Xie ◽

Todd C. Wehner ◽

Mark A. Conkling

Keyword(s):

Dna Sequence ◽

Restriction Enzyme ◽

Genomic Dna ◽

Sequence Variation ◽

Single Strand ◽

Sscp Analysis ◽

Enzyme Analysis ◽

Restriction Enzyme Analysis ◽

Pcr Products ◽

Dna Pcr

Combining the use of PCR and single-strand conformation polymorphisms (SSCP), nine sequences from the cucumber genome were successfully identified and cloned that encoded two well-conserved asparagine-proline-alanine (NPA) domain homologues to aquaporin genes. The sensitivity and detection efficiency of SSCP and restriction enzyme analysis for detecting DNA sequence variation were evaluated using similar-sized DNA fragments. The SSCP analysis was more sensitive and efficient for discriminating different clones than restriction enzyme analysis, although some sequence variation inside similar-sized DNA fragments could be identified by restriction analysis. Consideration of the results of SSCP analysis with DNA sequence information indicated that one or two base pair changes in the amplified regions could be detected. Moreover, the SSCP analysis results of genomic DNA PCR products that were amplified by degenerate primers can provide rough information about the number of member genes. If the SSCP bands of a cloned fragment (such as CRB7) did not have the corresponding bands from genomic DNA PCR products, that fragment might be a misamplified product. The PCR-based SSCP method with degenerate oligonucleotide primers should facilitate the cloning of member genes.

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Molecular cloning of Taenia solium genomic DNA and characterization of taeniid cestodes by DNA analysis

Parasitology ◽

10.1017/s003118200006683x ◽

1988 ◽

Vol 97 (1) ◽

pp. 161-176 ◽

Cited By ~ 26

Author(s):

A. K. Rishi ◽

D. P. McManus

Keyword(s):

Ribosomal Rna ◽

Restriction Enzyme ◽

Genomic Dna ◽

Taenia Solium ◽

Hybridization Analysis ◽

Enzyme Analysis ◽

Restriction Enzyme Analysis ◽

Restriction Enzyme Digestion ◽

Ribosomal Rna Gene ◽

Recombinant Plasmids

SUMMARYTotal DNAs, isolated from a range of taeniid cestodes (Taenia solium, T. saginata, T. pisiformis, T. crassiceps, T. hydatigena, T. ovis, T. multiceps and T. taeniaeformis), have been subjected to restriction enzyme digestion, Southern transfer and hybridization analysis using cloned fragments of the ribosomal RNA gene of Schistosoma mansoni. Substantial inter-specific genetic differences have been revealed on the basis of characteristic hybridization patterns for each of the taeniid cestode species. Furthermore, a random genomic DNA library has been constructed in the vector plasmid pAT153 using DNA extracted from a pig isolate (Indian origin) of T. solium. A panel of taeniid cestode DNAs including DNA from Echinococcus granulosus, has been used in conjunction with hybridization and restriction enzyme analysis to identify in the library a single recombinant plasmid with a T. solium-specific insert (coded pTS10) and two recombinant plasmids with T. solium inserts having selective specificities for T. solium and T. ovis (coded pTS17) and T. solium, T. saginata, T. ovis and T. multiceps (coded pTS28). These recombinant plasmids and the cloned fragments of the ribosomal RNA gene of S. mansoni have been used in restriction endonuclease, Southern transfer and hybridization analysis to detect intra-specific genetic variation in cysticerci of T. solium from India, Mexico and Zimbabwe. In addition, pTS10 and pTS17 have been used in a simple dot-blot assay to distinguish T. solium from T. saginata.

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