Role of HSP70 in cytoplasm protection against thermal stress in rohu, Labeo rohita.
To understand the function of HSP70 of Labeo rohita (LrHSP70) in cellular protection, LrHSP70 ORF cDNA was inserted into the plasmid of pET-32a(+) or pEGFP-L1. Then, the recombinant plasmids were transformed or transfected into Escherichia coli cells, mouse myeloma cells (MPC-11) or fish hepatoma cells (PLHC-1). Western blot results revealed that LrHSP70 was expressed in E. coli cells and molecular weight was estimated to be 70 kDa. In cells, LrHSP70 was over-expressed following thermal or cold stress. Results revealed that LrHSP70 protected prokaryotic cells against thermal or cold extremes as well as played the same role in MPC-11 and PLHC-1 cells. After heat treatment at 42 °C for 1 h, the viability of the cell was declined considerably. PLHC-1 cells with pEGFP-L1/LrHSP70 exhibited a higher survival rate (50%) than wild-type cells (18%) or cells with only pEGFP-L1 (21.2%). When the time lag extended to 2 h, the survival rates were 30%, 3.4% and 5.3% respectively. The present study revealed that LrHSP70 plays an important role in response to thermal and cold stress in fish.
['Giri|Sib Sankar|SS|', 'Sen|Shib Sankar|SS|', 'Sukumaran|V|V|']
D000818:Animals D015153:Blotting, Western D003530:Cyprinidae D003593:Cytoplasm D017931:DNA Primers D004591:Electrophoresis, Polyacrylamide Gel D005786:Gene Expression Regulation D018840:HSP70 Heat-Shock Proteins D006358:Hot Temperature D008099:Liver D051379:Mice D008807:Mice, Inbred BALB C D060888:Real-Time Polymerase Chain Reaction D020133:Reverse Transcriptase Polymerase Chain Reaction D013154:Spleen D013312:Stress, Physiological
Cytoplasmic protection Thermal stress Rohu Heat shock protein