Isolation Of A Putative Ap2/erf Transcription Factor Of Chinese Cabbage And Construction Of Its Expression Vector

AP2 transcription factors are important transcription factors which play very important roles in the adaption of plant to various stresses with significant functional diversity. To pursue function analysis of a AP2/ERF transcription factor in the process of heat shock response in Chinese Cabbage, a cDNA library of Chinese Cabbage leaves challenged with high temperature was screened with the specific primers of AP2/ERF proteins of Chinese cabbage on the basis of cDNA Library Screening method optimization. A putative AP2/ERF full length cDNA was acquired and its expression vector was constructed by Gateway cloning technology, The main results were as following:1. By the PCR Based 96-hole screening method, the cDNA library was screened with specific primers designed according to assemblying sequences with ESTs holomgy to a Arabidopsis thaliana AP2/ERF, a putative AP2/ERF positive clone was acquired, which was 812bp in length and harbored a open reading frame of 172 amino acids in length. By sequence aligment analysis, it was found that isolated cDNA clone shared different degrees homology with AP2 transcription factor of plants reported.2. By Gateway cloning technology, the putative Chinese cabbage AP2/ERF full length cDNA was cloned into enter vectors by BP reaction and then in destination vector, which could be used to transform dicotyledon by Agrobecteria mediaed method.Ther result of this thesis laid an important basis for the function identification of the putative AP2/ERF transcription factors in the heat shock reponse of Chinese Cabbage.