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Subtractive Cloning of DNA from Polymyxa graminis – an Obligate Parasitic Plasmodiophorid
Author(s) -
Šubr Z. W.,
Kastirr U.,
Kühne T.
Publication year - 2002
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1046/j.1439-0434.2002.00795.x
Subject(s) - biology , cloning (programming) , suppression subtractive hybridization , restriction enzyme , polymerase chain reaction , dna , restriction digest , molecular cloning , microbiology and biotechnology , plasmid , genomic dna , genetics , gene , complementary dna , cdna library , computer science , programming language
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 .