中间锦鸡儿CiNAC42基因序列分析及遗传转化

Sequence analysis and genetic transformation of CiNAC42 in Caragana intermedia

NAC转录因子是植物中最大的转录因子家族之一,在植物应答各种胁迫和生长发育过程中有重要的功能。该研究利用RACE技术克隆得到了CiNAC42基因的cDNA全长序列。生物信息学分析显示,该基因序列包括长度为933 bp的开放阅读框(ORF),可以编码311个氨基酸,N端具有保守的NAM结构域。利用实时荧光定量PCR对该基因的表达模式进行了检测,发现该基因在叶片中表达量最高,根中表达量最低。通过电极法将重组质粒pCanG-CiNAC42植物过表达载体转化到农杆菌GV3101中,利用浸花法转化野生型拟南芥(Arabidopsis thaliana),获得了阳性转基因拟南芥纯合子株系,为进一步研究转基因拟南芥功能提供了基础。

NAC transcription factor is one of the largest family of transcription factors in plants and plays an important role in plant response to various stresses and growth.In this study,the full-length cDNA fragments encoding NAC gene was amplified using RACE.Bioinformatics analysis showed that the CiNAC42 ORF was 933 bp,encoded 311 amino acids.The N-terminal had conservative NAM domain.Quantitative real-time PCR analysis revealed that the CiNAC42 expressed in different tissues,the expression was the highest in leaves and lowest in roots.The expression vector pCanG-CiNAC42 was constructed and transformed into Agrobacterium tumefaciens GV3101 by electrical shock method.The transgenic plants of Arabidopsis was obtained by Floral-dip method.And transgenic Arabidopsis homozygote strains were obtained,which provided the basis for further functional studies of transgenic Arabidopsis.