Analysis Of MeHsp70 Gene Function From Meloidogyne Enterolobii

During the research, the expression level of Hsp70 gene, as an important physiological index, often reflects the condition of the body.The Hsp70 gene will up-regulate to maintain the conformation of the key cellular proteins while the body encountering high temperature, hypoxia, cold or other harsh environment,in order to keep the cellular activities and reduce the damage to the body. With the infection of Meloidogyne enterolobii, the Hsp70 gene is mutating. Studying the related functions of M.enterolobii Hsp70 gene will lay foundations for controlling the Meloidogyne enterolobii disease.In the early, two prokaryotic expression vectors (pEASY-E1-MeHsp70 and pET30a-MeHsp70) that were transformed into Escherichia coli BL21, respectively, by the heat shock transformation way have been constructed successfully for the research.Keeping the original Escherichia coli BL21 and the transferred Escherichia coli BL21 into which the pEASY-E1-MeHsp70 recombinant plasmid, the pET30a-MeHsp70 recombinant plasmid and the no-load pEASY-Elwere transferred respectively in 55℃ or 65 ℃ for thermal stability of determination. The result shows that the transferred Escherichia coli BL21 of pET30a-MeHsp70 gene survive longer and have a better thermal stability than others.By the SDS-PAGE gel electrophoresis measurements, the MeHsp70 genes were all up-regulated expression, and the transferred Escherichia coli BL21 of pET30a-MeHsp70 gene owned higher protein content than others. By real-time fluorescence quantitative PCR test,Mesp70 gene does have a certain amount of up-regulated expression, which indicates that the increasing heat stability of Escherichia coli BL21 directly related to the up-regulation of MeHsp70 gene.Growth curves of the original Escherichia coli BL21 and the transferred Escherichia coli BL21 into which the pEASY-E1-MeHsp70 recombinant plasmid, the pET30a-MeHsp70 recombinant plasmid and the no-load pEASY-El were transferred respectively, were determined at 37 ℃ 200r/min and 30 ℃ and 200r/min. The growth curve test shows that the recombinant strain almost corrsponded with the original strain’s,and the strain in the pET30a-MeHsp70 vector was longer than the others.