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- W2225033504 abstract "The expression of a vast array of genes in plants changes and the physiological and metabolic reprogramming occurs against stress,which result in the enhanced stress resistance.Functional dissection of the differentially expressed genes is an approach to elucidate stress resistance mechanism.In order to acquire the full length cDNAs of the gene differentially expressed against Phytophthora capsici infection for functional genomics study of pepper disease resistance,a system including suppression subtractive hybridization cDNA library,normalized cDNA library was established in this study.The suppression subtractive hybridization cDNA library was constructed from pepper leaves inoculated with P.capsici,a number of SSH clones were randomly selected and sequenced.The results of BLASTN showed that 24 genes had homologous sequences in the database.These functionally known genes were detected to involve in energy metabolism,signal transduction,photosynthesis and stress resistance.The normalized cDNA library enriched in full-length sequences was constructed using DSN(duplex -specific nuclease)-normalization method combined with SMARTTM(switching mechanism at 5'end of RNA transcript) technique.The normalized cDNA library contained 1.8 × 106 independent clones and the average cDNA insert size was 1.75 kb with a recombination rate of 97%.The specific primers were designed according to the sequences of the differential expression ESTs from the SSH,and he corresponding cDNA clones were isolated from the normalized cDNA library by a PCR-based method.All the clones isolated from the cDNA library turned out to be full length cDNAs.The results indicated that the system could be used for isolation of full length cDNAs of genes differentially expressed against stresses such as P.capsici infection in pepper." @default.
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- W2225033504 date "2010-01-01" @default.
- W2225033504 modified "2023-09-27" @default.
- W2225033504 title "Establishment of full length cDNA isolation system of differentially expressed genes in Capsicum annuum against Phytophthora capsici infection and its application." @default.
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