梭梭14-3-3蛋白基因HaFT-9的克隆及特性分析

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

【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789bp的梭梭14-3-3蛋白基因,命名为HaFT-9。梭梭幼苗在20%PEG6000、200mmol/LNaCl和100μmol/LABA处理下,HaFT-9的表达量均在胁迫6h时达到峰值,在20μmol/LIAA处理下,HaFT-9的表达量在胁迫1h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。

【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.