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Cloning and gene expression of acetyl-CoA C-acetyl transferase gene
（AsAACT） from Aquilaria sinensis
LIU Juan XU Yan-hong YANG Yong LIANG Liang HAN Xiao-min GAO Zhi-hui ZHANG Zheng 2，
YANG Yun WEI Jian-he2*
（1.Institute of Medicinal Plant Development.Chinese Academy of Medical Sciences & Peking Union Medical College，
Beijing 100193， China； 2.Hainan Branch Institute of Medicinal Plant， Hainan Provincial Key Laboratory of Resources
Conservation and Development of Southern Medicine， Chinese Academy of Medical Sciences & Peking Union Medical College，
Wanning 571533， China； 3.Shandong University of Traditional Chinese Medicine， Jinan 250355， China；
4.Yanshan University， Qinhuangdao 066004， China）
[Abstract] Objective： This study aimed to clone the acetyl-CoA C-acetyl transferase （AACT） gene from Aquilaria sinensis and analyze the bioinformatics and expression of the gene. Method： One unique sequence containing partly AACT gene sequence was discovered in our previous transcriptome dataset of A. sinensis. AACT gene was cloned by RT-PCR and RACE strategy with the template of RNA extracted from A. sinensis stem. The bioinformatic analysis of this gene and its corresponding protein was performed. The AsAACT expression in calli was analyzed with GADPH gene as an internal control gene in wounded condition by qRT-PCR technique. Result： One unique sequence of AACT， named as AsAACT， was cloned from A. sinensis. The full length of AsAACT cDNA was containing a 1 236 bp ORF that encoded 411 amino acids. The result of qRT-PCR displayed that the highest expression level was at 4 h. which indicated that it was possibly involved in early-stage response to wound. Conclusion： Cloning and analyzing AsAACT gene from A. sinensis provided basic information for study the function and expression regulation of AsAACT in terpenoid biosynthesis.
[Key words] Aquilaria sinensis； AACT； MVA pathway； sequence analysis； expression analysis
doi：10.4268/cjcmm20140605