Subtractive Cloning of DNA from Polymyxa graminis- an Obligate Parasitic Plasmodiophorid

A simple subtractive hybridization was applied for cloning of Polymyxa graminis deoxyribonucleic acid (DNA). Total DNA preparations from concentrated P. graminis resting spores and from roots of non‐infested (healthy) barley were digested with different restriction enzymes and batch hybridized, foll...

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Bibliographic Details
Published in:Journal of phytopathology 2002-10, Vol.150 (10), p.564-568
Main Authors: Šubr, Z. W., Kastirr, U., Kühne, T.
Format: Article
Language:English
Subjects:
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biological and medical sciences
Biotechnology
DNA cloning
Fundamental and applied biological sciences. Psychology
Gene transfer
Genetic engineering
Genetic technics
Methods. Procedures. Technologies
Microbiology
polymerase chain reaction
Polymyxa graminis
subtractive hybridization
Synthetic digonucleotides and genes. Sequencing
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Summary:A simple subtractive hybridization was applied for cloning of Polymyxa graminis deoxyribonucleic acid (DNA). Total DNA preparations from concentrated P. graminis resting spores and from roots of non‐infested (healthy) barley were digested with different restriction enzymes and batch hybridized, followed by cloning in a plasmid vector. Sequencing and blast analysis of coincidentally selected clones enabled construction of polymerase chain reaction primers specific to P. graminis DNA. Four Polymyxa‐specific primers showed different affinities to DNA of type I and type II P. graminis and to DNA of P. betae.
ISSN:0931-1785
1439-0434
DOI:10.1046/j.1439-0434.2002.00795.x