Cloning and sequence analysis of DREB-like gene EeAP2.2 in Elytrigia elongata
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    Abstract:

    Due to the restriction of special nucleotide sequences in some genes, it’s difficult to obtain their full-length cDNA sequences by using only one method. In this paper, by combining RACE and Genome Walking techniques, a full-length cDNA sequence of AP2 family was cloned from Elytrigia elongata (2n=70), and named as EeAP2.2. Sequence analysis of EeAP2.2 revealed a single open reading frame (ORF) of 837 bp encoding a protein of 279 amino acids, and a highly conserved AP2 domain which classifies EeAP2.2 as a new member of AP2 superfamily. Further comparison analysis through NCBI blast showed that EeAP2.2 has 98%, amino sequence identities with TaDREB1 (Accession no. AAL01124.1) and TaDREBW50 (Accession no. AAY44605.1) from Triticum aestivum, and its amino sequence identities with HvDREB1-a (Accession no. AAY25517.1) from Hordeum vulgare, FaDREB2A (Accession no. CAG30547.1) from Festuca arundinacea and OsDREB2.2 (Accession no. AY064403) from Oryza sativa are 93%, 86%, 69%, respectively. The result indicates that EeAP2.2 has the highest homology with AP2 family members from Triticum aestivum. This study also provided a fast and effective method for cloning full length cDNA sequences in plants.

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History
  • Received:November 30,2010
  • Revised:February 28,2011
  • Adopted:May 23,2011
  • Online: August 15,2011
  • Published:
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