title: Role of HSP70 in cytoplasm protection against thermal stress in rohu, Labeo rohita.
abstract: 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.
authors.affiliation: Dept. of Biotechnology, Periyar Maniammai University, Thanjavur, 613403, Tamil Nadu, India. Electronic address: giribiotek@gmail.com.
authors.forename: Sib Sankar
authors.initials: SS
authors.lastname: Giri
authors.affiliation: Dept. of Pharmaceutical Technology, Jadavpur University, Kolkata, 700032, India. Electronic address: shibsankar.iicb@gmail.com.
authors.forename: Shib Sankar
authors.initials: SS
authors.lastname: Sen
authors.affiliation: Dept. of Biotechnology, Periyar Maniammai University, Thanjavur, 613403, Tamil Nadu, India.
authors.forename: V
authors.initials: V
authors.lastname: Sukumaran
mesh_terms: D000818:Animals; D015153:Blotting, Western; D003530:Cyprinidae / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation / Q000235:genetics / Q000502:physiology*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006358:Hot Temperature; D008099:Liver / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013154:Spleen / Q000378:metabolism; D013312:Stress, Physiological / Q000235:genetics / Q000502:physiology*
chemical_list: D017931:DNA Primers; D018840:HSP70 Heat-Shock Proteins
keywords: Cytoplasmic protection; Heat shock protein; Rohu; Thermal stress