珙桐CAC基因的克隆及其作为内参基因的评价

发布时间：2018-09-08 07:24

【摘要】：从珙桐(Davidia involucrata Baill.)中克隆到一个编码网格蛋白衔接蛋白复合物(Clathrin adaptor complexes,CAC)的基因,命名为DiCAC(Gen Bank登录号为KX268525)。该基因开放阅读框全长1 317 bp,编码438个氨基酸。同源序列比对与聚类分析表明,该基因与其它15种植物的CAC氨基酸序列的相似度达到94%以上,其中与葡萄CAC的氨基酸序列同源性最高。为评价该基因作为珙桐内参基因的可行性,结合5个珙桐管家基因(ACT7、EF1a、GAPDH、β-TUB、18S rRNA),采用半定量PCR和实时荧光定量PCR对DiCAC基因在珙桐不同组织以及不同发育阶段苞片中的表达稳定性进行分析,并与常见管家基因的稳定性进行比较,结果表明,该基因在珙桐不同组织以及不同发育阶段苞片中均为稳定表达,且表达稳定性优于常见管家基因,可以作为相关基因表达研究中的内参基因。首次克隆到珙桐CAC基因,并且明确了其作为内参基因的可行性,为深入研究珙桐相关基因的表达分析提供了候选内参基因资源。
[Abstract]:(Davidia involucrata Baill.) from Davidia involucrata A gene encoding the conjugate protein complex (Clathrin adaptor complexes,CAC) was cloned and named DiCAC (Gen Bank accession number (KX268525). The open reading frame of this gene encodes 438 amino acids with a total length of 1 317 bp,. The homologous sequence alignment and cluster analysis showed that the similarity of the CAC amino acid sequence between the gene and other 15 plants was over 94%, and the amino acid sequence homology with grape CAC was the highest. In order to evaluate the feasibility of this gene as an internal ginseng gene of Davidia involucrata, The expression and stability of DiCAC gene in bracts of different tissues and different developmental stages of Davidia involucrata were analyzed by semi-quantitative PCR and real-time fluorescence quantitative PCR combined with five housekeeping genes (ACT7,EF1a,GAPDH, 尾 -TUBX 18s rRNA), and the stability of DiCAC gene was compared with that of common housekeeper genes. The results showed that the gene was stably expressed in different tissues and bracts of Davidia involucrata, and its expression stability was superior to that of housekeeping genes. The CAC gene of Davidia involucrata was cloned for the first time, and its feasibility as an internal reference gene was confirmed, which provided candidate internal reference gene resources for further study on the expression and analysis of related genes of Davidia involucrata.
【作者单位】：中南林业科技大学生命科学与技术学院;
【基金】：中南林业科技大学博士后科研基金项目(0490016);中南林业科技大学青年基金项目(QJ201512)
【分类号】：S792.99;Q943.2

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