逆境胁迫下柽柳脂质转运蛋白基因 (ThLTP)的克隆与功能初步分析

林琳; 李健; 李慧玉; 穆怀志; 姜静

逆境胁迫下柽柳脂质转运蛋白基因 (ThLTP)的克隆与功能初步分析

1.
东北林业大学,林木遗传育种国家重点实验室,黑龙江哈尔滨 150040
基金项目:
转基因生物新品种培育科技重大专项(2009ZX08009-098B)
中图分类号: S793.5

Cloning and Function Analysis of Tamarix hispida Lipid Transfer Protein under Stress

1.
Northeast Forestry University,State Key Laboratory of Forest Genetics and Tree Breeding, Harbin 150040, Heilongjiang,China
CLC number: S793.5

摘要: 从以刚毛柽柳为材料构建的NaCl胁迫的根cDNA文库中测序得到1条LTP基因序列,其全长为635 bp、编码116个氨基酸,命名为ThLTP基因。采用实时荧光定量RT-PCR的方法,分别对0.2 mol·L-1 NaCl、150 μmol·L-1 CdCl2、20% (W/V) PEG6000、100 μmol·L-1 ABA及4 ℃下ThLTP基因在柽柳中的表达模式进行了分析,结果表明:ThLTP基因除了在4 ℃条件下根组织中为下调表达外,在其它处理中均表现为上调表达。将ThLTP基因克隆到大肠杆菌(Escherichia coli)的原核表达载体pET32a中,对重组菌Escherichia coli BL21(pET32a-LTP)的抗旱耐盐性进行分析。结果表明:在0.8% (W/V) NaCl和20% (W/V) PEG6000条件下,对照菌E. coli BL21(pET32a)无对数生长期,而重组菌E. colii BL21(pET32a-LTP)经过3 h的延迟生长后,进入到对数增长期,表明ThLTP可受盐、碱、低温诱导并能提高重组菌的抗旱耐盐能力。
- 刚毛柽柳
- / 脂质转移蛋白
- / 抗旱耐盐性
- / 表达分析
Abstract: In this study, a novel LTP gene which was named ThLTP was cloned from the cDNA library of Tamarix hispida root treated by 0.4 mol·L-1 NaCl. The sequence of ThLTP was 635 bp in full length, a protein with 116 amino acids residues. The authors examined the expression pattern of the ThLTP gene in leaf and root of T. hispida treated with 0.2 mol·L-1 NaCl, 150 μmol·L-1 CdCl2, 20% (W/V) PEG6000, 100 μmol·L-1 ABA and 4 ℃ stresses for different time using real time RT-PCR. The results showed that the ThLTP gene was induced by all the treatments in leaf and root except the treatment of 4 ℃ treated in root. ThLTP was inserted into a prokaryotic expression vector (pET32a) to produce the recombinant expression vector pET32a-LTP. The Escherichia coli BL21 (pET32a) could not live but the E. coli BL21 (pET32a-LTP) came to the maximum after 3 h delayed growth under the stress of 0.8% (W/V) NaCl and 20% (W/V) PEG6000. The results showed that ThLTP may be a salt- and drought-tolerant gene and enhance the stress tolerance of the recombinant.
- Tamarix hispida
- / LTP
- / salt- and drought-tolerant
- / expression analysis

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逆境胁迫下柽柳脂质转运蛋白基因 (ThLTP)的克隆与功能初步分析

Cloning and Function Analysis of Tamarix hispida Lipid Transfer Protein under Stress

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逆境胁迫下柽柳脂质转运蛋白基因 (ThLTP)的克隆与功能初步分析

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Cloning and Function Analysis of Tamarix hispida Lipid Transfer Protein under Stress

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逆境胁迫下柽柳脂质转运蛋白基因 (ThLTP)的克隆与功能初步分析

Cloning and Function Analysis of Tamarix hispida Lipid Transfer Protein under Stress

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逆境胁迫下柽柳脂质转运蛋白基因 (ThLTP)的克隆与功能初步分析

English Abstract

Cloning and Function Analysis of Tamarix hispida Lipid Transfer Protein under Stress

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