Cloning of a CAC Gene in Dove Tree（Davidia involucrata Baill.）and Evaluating its Potential as a Novel Reference Gene

doi:10.13560/j.cnki.biotech.bull.1985.2017.04.015

Abstract

Abstract: In this research,a gene named as DiCAC（GenBank accession number KX268525）,encoding the clathrin adaptor complexes（CAC）,has been cloned from dove tree（Davidia involucrata Baill.）. The sequence of DiCAC includes a ORF（open reading frame）of 1317 bp,which encodes a protein with 438 amino acids. Homologous alignment and cluster analysis show that DiCAC shared over 94% amino acid sequence identity with CAC genes in other 15 plants,and it has the highest amino acid homology with grape（Vitis vinifera）CACgene. To evaluate the feasibility of using DiCAC as a reference gene in dove tree,combined with five housekeeping genes of dove tree（ACT7,EF1a,GAPDH,β-TUB and 18S rRNA）,we analyzed the expression stability of DiCAC in different tissues and in bracts of different developmental stages. Furthermore,expression analysis of DiCAC detected by semi-quantitative PCR and qPCR showed that the expression levels of DiCAC are stable in both different tissues and bracts of different developmental stages. Remarkably,the expression of DiCAC showed more stability than the expression of common housekeeping genes. Therefore,it could be used as a reference gene of related gene expression research in dove tree. In this study,we have cloned the CAC gene from dove tree firstly and have confirmed the feasibility of its utilization as a reference gene. Our finding provides a novel candidate reference gene resources for the further research gene expression analysis in dove tree.

Key words: DiCAC, reference gene, bract, Dove tree（Davidia involucrata Baill.）

REN Rui, DAI Peng-hui, CAO Fu-xiang, DONG Xu-jie, LI Meng. Cloning of a CAC Gene in Dove Tree（Davidia involucrata Baill.）and Evaluating its Potential as a Novel Reference Gene[J]. Biotechnology Bulletin, 2017, 33(4): 119-129.

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