发菜耐旱相关蛋白NXL-01的基因克隆与表达分析
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国家自然科学基金(31360054;31060038)


Clone and Expression of Hypothetical Protein NXL-01 Related to Drought-tolerant in Nostoc flagelliforme
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    摘要:

    在发菜耐旱差异蛋白质组学研究中,发现假定蛋白(Hypothetical protein NXL-01)在干旱胁迫条件下表达量逐渐增加。根据已知氨基酸序列设计简并引物克隆NXL-01基因,长度为327 bp(GenBank登录号为HM854288)。生物信息学分析表明,该基因具有较高保守性,蛋白质二级结构主要由α螺旋和随机卷曲构成,是亲水性的膜外蛋白,有5个Ser磷酸化位点,1个Thr磷酸化位点。将NXL-01基因在大肠杆菌中表达,获得预期大小的重组蛋白(12.4 kD)。RT-PCR分析表明,NXL-01 mRNA在干旱胁迫条件下表达量逐渐增加,与NXL-01蛋白的表达趋势一致。

    Abstract:

    Nostoc flagelliforme is a kind of terrestrial nitrogen-fixing cyanobacteria being strong drought ecological adaptability.An unknown protein related to drought-tolerant,named hypothetical protein NXL-01,has been discovered by the proteomic analysis under drought stress.NXL-01 gene was cloned by designed degeneracy primers based on identified amino acid sequences.A full length of 327 bp DNA was obtained(GenBank access number:HM854288).Bioinformatics analysis showed that the NXL-01 gene was highly conservative.The secondary structure of NXL-01 was made up of α helix and random coil.It was a hydrophilic outside membrane protein with five Ser phosphorylation sites and one Thr phosphorylation site.NXL-01 was expressed in E.coli,and a 12.4 kD heterologous protein was observed.RT-PCR showed that mRNA level of NXL-01 gradually increased,consistent with the expression trend of NXL-01 in N.flagelliforme under drought stress.

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梁文裕,杨 佳,吴诗杰,等.发菜耐旱相关蛋白NXL-01的基因克隆与表达分析[J].西北植物学报,2015,35(1):44-49

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  • 在线发布日期: 2015-02-09
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