山葡萄转录因子CBF1基因的克隆及序列分析

生物技术通报 ›› 2013, Vol. 0 ›› Issue (8): 57-62.

山葡萄转录因子CBF1基因的克隆及序列分析

刘洋^1,2, 孙佳帝²

（1.吉林农业大学，长春 130118；2.长春职业技术学院，长春 130033）

收稿日期:2013-03-20 修回日期:2013-08-11 出版日期:2013-08-11 发布日期:2013-09-02
作者简介:刘洋，女，硕士，讲师，研究方向：植物基因工程；E-mail：274381295@qq.com
基金资助:
国家自然科学基金项目（30400300），吉林省杰出青年基金项目（20050117）

Cloning and Sequence Analysis of CBF1 Gene in Vitis amurensis

Liu Yang^1,2, Sun Jiadi²

（1. Agriculture University of Jilin, Changchun 130118；2. Changchun Vocational Institute of Technology, Changchun 130033）

Received:2013-03-20 Revised:2013-08-11 Published:2013-08-11 Online:2013-09-02

摘要/Abstract

摘要：

采用生物信息学方法比对多种低温诱导植物的CBF1同源基因，在其保守区域设计一对兼并引物，在特殊的RT-PCR条件下，结合反向PCR技术对山葡萄低温诱导转录因子CBF1的全长基因进行了克隆，经测序和序列分析显示，山葡萄转录因子CBF1基因ORF长为753 bp，无内含子，编码251个氨基酸，编码蛋白分子量和等电点分别为27.503 kD和3.11，属酸性蛋白质。应用Blast软件将山葡萄转录因子CBF1基因与大麦(Hordeum vulquare)、水稻(Oryza sativa)等10种植物CBF1基因氨基酸序列比较同源性达到80.7%，且在AP2/EREBP结构域有高度的保守性，表明已成功克隆山葡萄(Vitis amurensis)转录因子CBF1基因，将其命名为ViCBF1, 在GenBank上的登录号为DQ517296。

关键词: 山葡萄(Vitis amurensis), 低温诱导, 转录因子, CBF1基因, COR基因, 克隆

Abstract:

Bioinformatic method was used for the comparative analysis of CBF1 gene sequences in cold acclimation, conservative domains among the sequences were determined for degenerate primer designing, the defined PCR conditions and Reverse PCR clone were adopted to clone the full length CBF1 genes from genes of Vitis amurensis which is in cold acclimation. Sequence analysis showed that the ORF lengths of CBF1 gene is 753 bp and has no intron in coding domain, and encoded proteins of 251 residues. Furthermore, their molecular weights and isoelectric points of 27.503 kD and 3.11 were predicted from the calculated pI values. Similarity of amino acids to many plant which is Hordeum vulquare and Oryza sativa is 80.7%, it have a highly conserved sequence in AP2/EREBP, this demonstrate we clone transcription factor CBF1 genes of Vitis amurensis . It is named ViCBF1 which is registered in GenBank, number is DQ517296.

Key words: Wild grape(Vitis amurensis), Cold acclimation, Transcription factor, CBF1 gene, COR gene, Cloning

刘洋, 孙佳帝. 山葡萄转录因子CBF1基因的克隆及序列分析[J]. 生物技术通报, 2013, 0(8): 57-62.

Liu Yang, Sun Jiadi. Cloning and Sequence Analysis of CBF1 Gene in Vitis amurensis[J]. Biotechnology Bulletin, 2013, 0(8): 57-62.

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