胁迫诱导型启动子rd29A的克隆及其序列分析

Cloning and Sequence Analysis of the Stress-induced Promoter rd29A

采用CTAB法提取拟南芥(Arabidopsis thaliana)叶片总DNA,设计并合成一对引物,通过PCR扩增得到一特异片段,并连接至pGEM-T Easy Vector上进行克隆测序。结果表明该片段全长为744 bp,与报道序列(AY973635)存在三个碱基的差异,同源性达99.6%;且该片段含有1个TATA box(TATAAA)、1个CAAT box(GCCAAT)、4个干旱、高盐和低温响应DRE(dehydration responsive element binding protein)顺式作用元件(核心序列为CCGAC),从而为后期胁迫诱导型植物表达载体的构建及遗传转化奠定了基础。

Genomic DNA was isolated from Arabidopsis thaliana by CTAB method.A pair of specific primers was designed and synthesized according to the reported sequence of the stress-induced promoter rd29A in GenBank.The PCR product was amplified, ligated into pGEM-T Easy Vector and sequenced.The results showed that the specific fragment was 744 bp;that there were three different bases between the sequence tested in this paper and the one reported in GenBank(AY973635),whose homology in nucleotide sequences reached 99.6%,that it contained one TATA box,one CAAT box and four DRE cis-action elements responsive to drought, high salt and low temperature;which laid the foundation for the future part of its construction of plant expression vector and genetic transformation.