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Indian J Biochem Biophys ; 2014 Aug; 51(4): 302-307
Article Dans Anglais | IMSEAR | ID: sea-154252

Résumé

Full-length cDNAs are very important for genome annotation and functional analysis of genes. The number of full-length cDNAs from watermelon remains limited. Here we report first the construction of a full-length enriched cDNA library from Fusarium wilt stressed watermelon (Citrullus lanatus Thunb.) cultivar PI296341 root tissues using the SMART method. The titer of primary cDNA library and amplified library was 2.21 × 106 and 2.0 × 1010 pfu/ml, respectively and the rate of recombinant was above 85%. The size of insert fragment ranged from 0.3 to 2.0 kb. In this study, we first cloned a gene named ClWRKY1, which was 1981 bp long and encoded a protein consisting of 394 amino acids. It contained two characteristic WRKY domains and two zinc finger motifs. Quantitative real-time PCR showed that ClWRKY1 expression levels reached maximum level at 12 h after inoculation with Fusarium oxysporum f. sp. niveum. The full-length cDNA library of watermelon root tissues is not only essential for the cloning of genes which are known, but also an initial key for the screening and cloning of new genes that might be involved in resistance to Fusarium wilt.


Sujets)
Séquence d'acides aminés , Séquence nucléotidique , Citrullus/génétique , Amorces ADN , ADN complémentaire/génétique , Fusarium/pathogénicité , Gènes de plante , Données de séquences moléculaires , ARN des plantes/génétique , Réaction de polymérisation en chaine en temps réel , Recombinaison génétique , Similitude de séquences d'acides aminés , Facteurs de transcription/composition chimique , Facteurs de transcription/génétique
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