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PCR-based cloning of two Ξ-1,4-endoglucanases from the root-lesion nematode Pratylenchus penetrans

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Polymerase chain reaction (PCR) with degenerate primers was performed on cDNA from Pratylenchus penetrans to amplify a conserved region in β-1,4-endoglucanase genes. Two distinct genomic DNA fragments that encoded partial β-1,4-endoglucanase sequences were isolated. Two corresponding full-length cDNAs, designated Pp-eng-1 and Pp-eng-2, were obtained by 3′ and 5′ rapid amplification of cDNA ends. Pp-eng-1 and Pp-eng-2 consisted of 1521 nucleotides encoding 455 amino acids and 1179 nucleotides encoding 328 amino acids, respectively. The two cloned genes encoded family 5 β-1,4-endoglucanases. The predicted protein PP-ENG-1 contained a cellulose-binding domain (CBD), and the predicted protein PP-ENG-2 lacked a CBD.


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Affiliations: 1: Laboratory Plant Nematology, Hokkaido National Agricultural Experimental Station, Hitujigaoka-1, Toyohira, Sapporo 062-8555, Japan


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