摘要
[目的]探索CBF转录激活因子基因在侧金盏花中是否存在。[方法]以侧金盏花基因组DNA为模板,根据拟南芥CBF基因序列的保守区设计1对特异性引物,通过PCR扩增获得1个DNA片段,将其克隆到pUCm-T载体中,转化大肠杆菌TG1,筛选阳性克隆并对其进行测序鉴定。[结果]测序结果显示通过PCR扩增获得的基因片段长448 bp。在GenBank中检索,该序列与从拟南芥和其他作物中克隆的CBF同源基因序列同源性较低,说明是一个新的基因片段,初步认定该片段是侧金盏花CBF基因片段。[结论]该研究为进一步克隆侧金盏花CBF转录激活因子基因全长序列、鉴定其生物学意义奠定了基础。
[Objective] The aim was to explore whether the CBF transcriptional activator gene existed in Adonis amurensis.[Method] With the genomic DNA of A.amurensis as template,a DNA fragment was obtained by PCR amplification using 1 pair of specific primers designed from conservative region of Arabidopsis thaliana CBF gene sequence.The fragment was cloned into pUCm-T vector,then transformed into Escherichia coli TG1,and then sequenced and identified through screening the positive clones.[Result]The sequencing result showed that a fragment with the length of 448 bp was obtained by PCR amplification.The sequence shared lower homology with the CBF homologous gene sequences cloned from A.thaliana and other crops,which showed that it was a new gene fragment and the fragment was identified as CBF gene fragment from A.amurensis preliminarily.[Conclusion] The research laid the foundation for the further study of the clone of CBF transcriptional activator gene with full length sequence from A.amurensis and the identification of its biological significance.
出处
《安徽农业科学》
CAS
北大核心
2008年第27期11686-11687,共2页
Journal of Anhui Agricultural Sciences
基金
河南省教育厅自然科学基金项目(2007180028)
河南师范大学青年科学基金项目(2006037)
