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当前位置: 首页 > 文章 > 香樟Actin基因的克隆及表达分析 生物技术通报 2015,31 (5) 120-127
Position: Home > Articles > Cloning and Expression Analysis of Actin Gene in Cinnamomum camphora Biotechnology Bulletin 2015,31 (5) 120-127

香樟Actin基因的克隆及表达分析

作  者：
李勇鹏;张力维;姚瑶;黄蕊;杜丽
单  位：
南阳师范学院生命科学与技术学院
关键词：
香樟;Actin;基因克隆;表达分析;实时定量PCR
摘  要：
Actin作为一个看家基因,经常在实时定量PCR中被用作内参对不同样品中的m RNA进行定量,因此在基因表达分析中扮演重要的角色。利用同源克隆的方法,根据Gen Bank中已经公布的其他植物的Actin基因的保守序列设计简并引物,采用RT-PCR(Reverse Transcription Polymerase Chain Reaction)技术从香樟中获得了4个c DNA片段。分子生物学分析软件分析结果显示,4个片段大小为均为998 bp,编码332个氨基酸;同源性分析表明,所获得的片段属于Actin亚家族,分别命名为Cc ACTa、Cc ACTb、Cc ACTc和Cc ACTd并提交Gen Bank(登录号:KM086736、KM086737、KM086738和KM086739)。实时定量PCR结果显示,Cc ACTc在根、茎、叶及低温处理的叶片中表达量相对稳定,可以作为候选内参基因。
译  名：
Cloning and Expression Analysis of Actin Gene in Cinnamomum camphora
作  者：
Li Yongpeng;Zhang Liwei;Yao Yao;Huang Rui;Du Li;School of Life Science and Technology,Nangyang Normal University;
关键词：
Cinnamomum camphora;;Actin;;gene cloing;;expression analysis;;quantitative real-time PCR
摘  要：
As a house-keeping gene, Actin has been always being used as an internal standard to normalize m RNA levels between different samples by quantitative real-time PCR, thus plays an important role in gene expression analysis. In this research, through a method of homology cloning, a pair of degenerate primers were designed based on the conserved sequences of Actin genes from other plants submitted to Gen Bank, and then 4 c DNA fragments from Cinnamomum camphora were obtained using RT-PCR. Molecular biological analysis showed that, each of the 4 c DNA fragments was 998 bp and encoded a putative protein of 332 amino acids. Moreover, according to the homology analysis, the 4 c DNA fragments belonged to Actin subfamily, and they were named Cc ACTa, Cc ACTb, Cc ACTc and Cc ACTd, and deposited in Gen Bank(Accession number :KM086736 KM086737 KM086738 and KM086739). Quantitative real-time PCR results revealed that the expression level of Cc ACTc in different organs such as root, stem, leaf and leaves under low temperature treaments was relatively stable. It could serve as a candidate reference gene.

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