| Expression in Escherichia coli of Three Different Soybean Late Embryogenesis Abundant （LEA） Genes to Investigate Enhanced Stress Tolerance |
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| 作者姓名： | YingLAN DanCAI Yi-ZhiZHENG |
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| 作者单位： | [1]InstituteofGeneticsandCytology,SchoolofLifeSciences,NortheastNormalUniversity,Changchun130021,China／／MedicalCollegeofBeihuaUniversity,Jilin132021,China [2]CollegeofLifeSciences,KeyLaboratoryofMicroorganismandGeneticEngineeringofShenzhenCity,ShenzhenUniversity,Shenzhen518060,China |
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| 摘 要： | In order to identify the function of late embryogenesis abundant (LEA) genes, in vitro functional analyses were performed using an Escherichia coli heterologous expression system. Three soybean late embryogenesis abundant (LEA) genes, PMll (GenBank accession No. AF004805; group 1), PM30(AF117884; group 3), and ZLDE-2 (AY351918; group 2), were cloned and expressed in a pET-28a system.The gene products were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified by mass spectrometry. E. coli cells containing the recombinant plasmids or empty vector as controls were treated by salt and low temperature stress. Compared with control cells, the E. coli cells expressing either PMll or PM30 showed a shorter lag period and improved growth when transferred to LB (Luria-Bertani) liquid media containing 800 mmol/L NaC1 or 700 mmol/L KC1 or after 4℃ treatment. E. coli cells expressing ZLDE-2 did not show obvious growth improvement both in either high KC1 medium or after 4℃ treatment. The results indicate that the E. coli expression system is a simple, useful method to identify the functions of some stress-tolerant genes from plants.
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| 关 键 词： | 大肠杆菌 大豆 抗逆性 胚胎形成后期 基因表达 |
Expression in Escherichia coli of Three Different Soybean Late Embryogenesis Abundant (LEA) Genes to Investigate Enhanced Stress Tolerance |
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| Authors: | Ying LAN Dan CAI Yi-Zhi ZHENG |
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| Institution: | Institute of Genetics and Cytology, School of Life Sciences, Northeast Normal University, Changchun 130021, China;;College of Life Sciences, Key Laboratory of Microorganism and Genetic Engineering of Shenzhen City, Shenzhen University, Shenzhen 518060, China;;Medical College of Beihua University, Jilin 132021, China |
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| Abstract: | Abstract: In order to identify the function of late embryogenesis abundant (LEA) genes, in vitro functional analyses were performed using an Escherichia coli heterologous expression system. Three soybean late embryogenesis abundant (LEA) genes, PM11 (GenBank accession No. AF004805; group 1), PM30 (AF117884; group 3), and ZLDE-2 (AY351918; group 2), were cloned and expressed in a pET-28a system. The gene products were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified by mass spectrometry. E. coli cells containing the recombinant plasmids or empty vector as controls were treated by salt and low temperature stress. Compared with control cells, the E. coli cells expressing either PM11 or PM30 showed a shorter lag period and improved growth when transferred to LB (Luria-Bertani) liquid media containing 800 mmol/L NaCl or 700 mmol/L KCl or after 4°C treatment. E. coli cells expressing ZLDE-2 did not show obvious growth improvement both in either high KCl medium or after 4°C treatment. The results indicate that the E. coli expression system is a simple, useful method to identify the functions of some stress-tolerant genes from plants.
(Managing editor: Li-Hui ZHAO) |
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| Keywords: | Escherichia coli gene expression LEA gene osmotic stress salt stress soybean |
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