扁桃CBF1基因的克隆与生物信息学分析

doi:10.11924/j.issn.1000-6850.2014-1877

中国农学通报 ›› 2014, Vol. 30 ›› Issue (34): 16-23.doi: 10.11924/j.issn.1000-6850.2014-1877

所属专题：生物技术；园艺

扁桃CBF1基因的克隆与生物信息学分析

张亮¹，张智俊²，李疆³，代培红¹，田嘉³，罗淑萍¹

（¹新疆农业大学农学院，乌鲁木齐 830052；²浙江农林大学亚热带森林培育国家重点实验室培育基地，
浙江临安 311300；³新疆农业大学林学与园艺学院，乌鲁木齐 830052）

收稿日期:2014-07-07 修回日期:2014-11-23 接受日期:2014-09-30 出版日期:2015-03-10 发布日期:2015-03-10
通讯作者: 张亮
基金资助:

基金项目：国家自然科学基金“新疆扁桃耐寒基因CBF1在花粉发育中的功能及其花粉发育避寒机制研究”(31360473)；国家自然科学基金“新疆野扁桃不同逆境梯度下的繁殖对策与生态适应性研究”(31260186)；新疆自治区“十二五”重大科技专项“新疆特色果树种质资源利用技术研究与示范”(201130102-1)；新疆自治区果树学重点学科基金。

Cloning and Bioinformatic Analysis of CBF1 Gene in Amygdalus communis

Zhang Liang¹, Zhang Zhijun², Li Jiang³, Dai Peihong¹, Tian Jia³, Luo Shuping¹

(¹College of Agronomy, Xinjiang Agricultural University, Urumqi 830052;
²The Nurturing Station for the State Key Laboratory of Subtropical Silviculture,
Zhejiang A&F University, Lin’an Zhejiang 311300;
³College of Forestry and Horticulture, Xinjiang Agricultural University, Urumqi 830052)

Received:2014-07-07 Revised:2014-11-23 Accepted:2014-09-30 Online:2015-03-10 Published:2015-03-10

摘要/Abstract

摘要： 为揭示CBF基因在扁桃抗逆机制中发挥的功能，对扁桃CBF1基因进行克隆与分析。以新疆扁桃主栽品种‘鹰嘴’为材料，根据已经报道的CBF1基因序列设计引物，通过PCR技术从扁桃基因组DNA中获得CBF1基因，命名为YZ-AcCBF1，并已登录GenBank（登录号KJ818900）。经生物信息学分析，该基因开放阅读框为729 bp，编码242个氨基酸，分子式为C₁₁₉₅H₁₈₈₆N₃₄₆O₃₆₈S₁₃，相对分子质量为27.405 kDa，等电点为7.69，半衰期为30 h，脂肪系数为63.72，疏水性平均值为-0.646。对其亚细胞定位、跨膜结构、信号肽及疏水性/亲水性进行分析，表明其定位在细胞核，不存在跨膜结构，为非分泌性亲水蛋白。系统发育分析结果显示其与桃(AGS13699)的亲缘关系最近。二级结构预测显示该蛋白主要由26.86%的α螺旋和58.68%的无规卷曲组成。三维结构预测表明其符合AP2结构域模型特点。

关键词: 噁唑酰草胺, 噁唑酰草胺, 光解, 半衰期

Abstract: The AcCBF1 gene was cloned and analyzed in order to identify the role of CBF transcription factors in plant resistance of Amygdalus communis. The CBF1 homologue gene named YZ- AcCBF1 (Genbank Accession No. KJ818900) was isolated from‘Yingzui’(Amygdalus communis) with the technology and method of PCR. The bioinformatics analysis showed that the open reading frame was 729 bp in full- length and encodedaprotein of 242 amino acids. The molecular formula was C1195H1886N346O368S13, and its relative molecular mass was approximately 27.405 kDa and isoelectric point was 7.69. This protein was an instable protein whose deduced half-life was 30 h. Its fat factor was 63.72 and the average of hydrophobic was -0.646. Subcellular localization analysis showed that AcCBF1 protein was localized in the nucleus, it was not a secreted protein but the hydrophilic protein and the transmembrane domain did not exist. Phylogenetic analysis revealed that AcCBF1 encode protein showed the highest similarity with Prunus persica (AGS13699). The prediction of secondary structure of protein indicated that the encoded protein mainly contain 26.86% ofα- helices and 58.68% of the random coil. Three- dimensional structure predicted that AcCBF1 protein confirmed the characteristics of AP2 domain model.

张亮,张智俊,李疆,代培红,罗淑萍. 扁桃CBF1基因的克隆与生物信息学分析[J]. 中国农学通报, 2014, 30(34): 16-23.

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扁桃CBF1基因的克隆与生物信息学分析

Cloning and Bioinformatic Analysis of CBF1 Gene in Amygdalus communis

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