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 embryoge...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.展开更多
基金the National Special Program for Research and Industrialization of Transgenic Plants，国家自然科学基金，Foundation for Key Teachers in Universities
文摘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.
