Arak University Cell and Tissue Journal 2228-7035 8 1 2017 03 21 Study the effects of some acute environmental conditions on recombinant Hsp70 protein expression in Rutilus Frissi Kutum liver in E. coli Study the effects of some acute environmental conditions on recombinant Hsp70 protein expression in Rutilus Frissi Kutum liver in E. coli 32 41 26496 10.52547/JCT.8.1.32 FA O Saberi Department of Biology, University of Guilan, University Campus 2, Rasht, Iran H Ghafoor Department of Biology, Faculty of science, University of Guilan, Rasht, Iran Journal Article 2017 03 21 Aim: The aim of the present study was to investigate the optimal expression of Hsp70 recombinant Rutilus FrissiKutum liver in E. coli. Material and methods: To evaluating of expression level of recombinant Hsp70, transformed E. coli were cultured in LB medium at 2, 4, 8, 12, 24, 36 and 48 hours, 20, 25, 30, 37 and 50 °C, extreme pH (5 and 9) and in present of heavy metals e.g. mercury, cobalt, iron, zinc and etc. Expression of recombinant Hsp70 protein was determined by SDS-PAGE 12% analysis of E. coli extracts followed by staining with Coomassie Blue. Results: Optimum expression of recombinant Hsp70 was obtained at 4 and 8 h, 37 °C and pH 5. Additionally, the maximum and minimum expression of this protein was achieved in present of manganese and cobalt respectively. The results further indicate that the survival of E. coli with Hsp70 was enhanced compared to the control cells. Conclusion: All results reveal that transformed bacteria are able to survive against extreme environmental conditions compared to control sample, because of having recombinant Hsp70. This heat-shock protein protects vital proteins from denaturing by its chaperone property. So, growth condition described in this study can be used for optimizing production of recombinant Hsp70 in E. coli host. Aim: The aim of the present study was to investigate the optimal expression of Hsp70 recombinant Rutilus FrissiKutum liver in E. coli. Material and methods: To evaluating of expression level of recombinant Hsp70, transformed E. coli were cultured in LB medium at 2, 4, 8, 12, 24, 36 and 48 hours, 20, 25, 30, 37 and 50 °C, extreme pH (5 and 9) and in present of heavy metals e.g. mercury, cobalt, iron, zinc and etc. Expression of recombinant Hsp70 protein was determined by SDS-PAGE 12% analysis of E. coli extracts followed by staining with Coomassie Blue. Results: Optimum expression of recombinant Hsp70 was obtained at 4 and 8 h, 37 °C and pH 5. Additionally, the maximum and minimum expression of this protein was achieved in present of manganese and cobalt respectively. The results further indicate that the survival of E. coli with Hsp70 was enhanced compared to the control cells. Conclusion: All results reveal that transformed bacteria are able to survive against extreme environmental conditions compared to control sample, because of having recombinant Hsp70. This heat-shock protein protects vital proteins from denaturing by its chaperone property. So, growth condition described in this study can be used for optimizing production of recombinant Hsp70 in E. coli host.

https://jct.araku.ac.ir/article_26496_1697fe5eb0141dca1379090a4d766cc7.pdf