| 刺山柑70 kD热休克蛋白基因克隆及原核表达 |
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| 引用本文： | 栾东东,董玉芝. 刺山柑70 kD热休克蛋白基因克隆及原核表达[J]. 西北植物学报, 2009, 29(7) |
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| 作者姓名： | 栾东东 董玉芝 |
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| 作者单位： | 1. 新疆农业大学,林学与园艺学院,乌鲁木齐,830052
2. 新疆农业大学,林学与园艺学院,乌鲁木齐,830052;新疆农业大学,农业生物技术重点实验室,乌鲁木齐,830052 |
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| 基金项目： | 国家环保局环境保护项目 |
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| 摘 要： | 以干旱区特有的抗逆性植物刺山柑为材料,利用RT-PCR结合cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)方法,克隆了一个HSP70基因,将该基因进行原核表达,通过对大肠杆菌进行温度胁迫实验,并计算存活率来验证该基因对细胞的保护作用.结果表明:该基因全长2 202 bp,开放阅读框共1 950 bp,编码649个氨基酸,表达产物分子量为71.044 6 kD;序列分析表明,该基因属于HSP70基因家族,将该基因命名为CsHSP70,并提交到GenBank,登录号为EU574936.构建了CsHSP70基因的原核表达载体,并使重组质粒pGEX-4T-2-CsHSP70在大肠杆菌中异源表达,对大肠杆菌进行温度胁迫实验结果显示,在高温(50℃)和低温(4℃)条件下,转重组质粒的大肠杆菌在两种胁迫下存活率均高于对照,说明在温度胁迫下CsHSP70对大肠杆菌细胞具有保护作用.
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| 关 键 词： | 刺山柑 热休克蛋白 原核表达 |
Cloning and Its Prokaryotic Expression of a 70 kD Heat Shock Protein Gene from Capparis spinosa |
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| LUAN Dong-dong,DONG Yu-zhi. Cloning and Its Prokaryotic Expression of a 70 kD Heat Shock Protein Gene from Capparis spinosa[J]. Acta Botanica Boreali-Occidentalia Sinica, 2009, 29(7) |
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| Authors: | LUAN Dong-dong DONG Yu-zhi |
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| Affiliation: | 1 College of Forestry and Horticulture;Xinjiang Agricultural University;Urumqi 830052;China;2 Key Laboratory of Agrobiotechnology;China |
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| Abstract: | By the methods of RT-PCR and rapid amplification of cDNA ends(RACE) we isolated a HSP70 gene(CsHSP70) from Capparis spinosa,a specific resistant plant in arid zone.Recombinant CsHSP70 was overexpressed in Escherichia coli and verify that the gene's protective effect on cells by the temperature treatment experiment.The results showed that the CsHSP70 gene contained 2 202 nucleotides with a 1 950 nucleotides length open reading frame(ORF) coding 649 amino acid residues with the predicted molecular weight of 7... |
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| Keywords: | RACE Capparis spinosa heat shock protein rapid amplification of cDNA ends prokaryotic expression |
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