Cloning and Characterization Analysis of HaFT-9 Gene in14-3-3 Protein in Haloxylon ammodendron

doi:10.6048/j.issn.1001-4330.2019.06.011

Abstract

Abstract: 【Objective】 To analyze the expression pattern of Haloxylon ammodendron 14-3-3 protein gene HaFT-9 under different stresses, and the binding characteristics of HaFT-9 protein, so as to provide a theoretical basis for studying the molecular mechanism of stress resistance in Haloxylon ammodendron.【Method】A 14-3-3 protein gene, HaFT-9, was cloned using Haloxylon ammodendron cDNA as a template, and the expression pattern of this gene under different stresses was analyzed by qRT-PCR, and the HaFT-9 protein binding properties were verified by yeast two-hybrid assay.【Result】The gene for the Haloxylon ammodendron 14-3-3 protein with an ORF length of 789 bp was cloned and named HaFT-9. Under 20% PEG6000, 200 mmol/L NaCl, and 100 μmol/L ABA treatment, the expression of HaFT-9 all peaked at 6 h of stress, and under 20 μmol/L IAA treatment, the expression of HaFT-9 peaked at 1 h of stress. HaFT was up-regulated in the normal temperature recovery stage in the adaptation experiment of high temperature stress by simulating surface high temperature stress at 40℃, 55℃and 55℃, and 65℃. Yeast two-hybrid experiment showed that HaFT-9 could form homologous dimer with itself.【Conclusion】The HaFT-9 gene (accession number API85525.1) was cloned, which could respond to drought stress, salt stress, ABA and IAA treatment, and was associated with adaptability to high temperature stress. HaFT-9 protein can interact with itself to form homodimer.

Key words: 14-3-3 protein; Haloxylon ammodendron; clone; Real-time fluorescence quantification; yeast two-hybrid

摘要： 【目的】分析梭梭14-3-3蛋白基因HaFT-9在不同胁迫下的表达模式,解析HaFT-9蛋白结合特性,为研究梭梭抗逆分子机制提供理论基础。【方法】以梭梭cDNA为模板克隆14-3-3蛋白基因HaFT-9,利用qRT-PCR技术对该基因在不同胁迫下的表达模式进行分析,并通过酵母双杂交实验验证HaFT-9蛋白结合特性。【结果】克隆得到ORF长度为789 bp的梭梭14-3-3蛋白基因,命名为HaFT-9。梭梭幼苗在20% PEG6000、200 mmol/L NaCl和100 μmol/L ABA处理下,HaFT-9的表达量均在胁迫6 h时达到峰值,在20 μmol/L IAA处理下,HaFT-9的表达量在胁迫1 h时达到峰值。梭梭幼苗经40、55℃和55、65℃模拟地表高温胁迫,高温胁迫适应性试验中的常温复原阶段HaFT-9均上调表达。酵母双杂交实验表明HaFT-9可与自身形成同源二聚体。【结论】克隆得到HaFT-9基因(登录号为API85525.1),该基因可响应干旱胁迫、盐胁迫、ABA和IAA处理,并与高温胁迫适应性相关,HaFT-9蛋白能与自身互作形成同源二聚体。

关键词: 14-3-3蛋白, 梭梭, 克隆, 实时荧光定量, 酵母双杂

CLC Number:

Q786

LIU Shuan-shuan, WANG Bo, ZONG Xing-feng, WANG Jiao, WANG Xin-qi, YAO Zheng-pei, REN Yan-ping, ZHANG Hua. Cloning and Characterization Analysis of HaFT-9 Gene in14-3-3 Protein in Haloxylon ammodendron [J]. Xinjiang Agricultural Sciences, 2019, 56(6): 1083-1094.

刘栓栓, 王波, 宗兴风, 王娇, 王新奇, 姚正培, 任燕萍, 张桦. 梭梭14-3-3蛋白基因HaFT-9的克隆及特性分析[J]. 新疆农业科学, 2019, 56(6): 1083-1094.

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