Expression of heat shock protein70-1 gene in bovine peripheral blood mononuclear cells

DOI: 10.5958/0976-0555.2015.00068.0    | Article Id: B-2620 | Page : 325-327
Citation :- Expression of heat shock protein70-1 gene in bovine peripheral blood mononuclear cells.Indian Journal Of Animal Research.2015.(49):325-327
Mehtab Singh Parmar*, A.K. Madan, S.K. Rastogi and Ruokuobeinuo Huozha mehtab.parmar@gmail.com
Address : Department of Veterinary Physiology and Biochemistry, College of Veterinary and Animal Sciences, Pantnagar-263 145, India.

Abstract

The present study has demonstrated the expression of HSP70-1 gene in bovine peripheral blood mononuclear cells (PBMCs). Blood samples were collected aseptically by jugular vein puncture in EDTA (1mg/ml) from six apparently healthy sahiwal cows (Bos Indicus). Peripheral blood mononuclear cells (PBMCs) were separated from the blood using HiSepTM Lymphocyte Separation Media (LSM). Total RNA was isolated from PBMCs using Trizol reagent. The purity and concentration of total RNA was checked using Nanodrop instrument whose further confirmation was made by using denaturing agarose gel (1.5%) electrophoresis. Isolated total RNA was reverse transcribed to cDNA that was used for amplification of HSP70-1 gene. Full length open reading frame (ORF) of HSP70-1 gene was amplified using specific primers pair. The presence of 1926 base pair band in denaturing agarose indicated the amplification of HSP70-1 gene. To conclude, our results demonstrate that HSP70-1 gene is expressed in bovine PBMCs.

Keywords

HSP70 gene Peripheral blood mononuclear Cells Sahiwal cows.

References

  1. Beckham, J.T.; Mackanos, M.A.; Crooke, C.; Takahashi, T.; O’Connell-Rodwell, C.; Contag, C.H. and Jansen, E.D. (2004). Assessment of cellular response to thermal laser injury through bioluminescence imaging of heat shock protein 70. Photochem Photobiol. 79:76–85.
  2. Christians, E.; Michel, E.; Adenot, P.; Mezger, V.; Rallu, M.; Morange, M. and Renard, J.P. (1997). Evidence for the involvement of mouse heat shock factor 1 in the atypical expression of the HSP70.1 Heat shock gene during mouse zygotic genome activation. Mol Cell Biol. 17:778–788.
  3. Dangi, S.S.; Gupta, M.; Maurya, D.; Yadav, V.P.; Panda, R.P.; Singh, G.; Mohan, N.H.; Bhure, S.K.; Das, B.C.; Bag, S.; Mahapatra, R.; Sharma G.T. and Sarkar, M. (2012). Expression Profile of HSP Genes during Different Seasons in Goats (Capra hircus). Trop Anim Health Pro. 12:155-158.
  4. Fargnoli, J.; Kunisada, T.; Fornace, A.J. Jr.; Schneider, E.L. and Holbrook, N.J. (1990). Decreased expression of heat shock protein 70 mRNA and protein after heat treatment in cells of aged rats. Proc. Natl. acad. Sci. 87: 846-850.
  5. Gade, N.; Mahapatra, R. K.; Sonawane, A.; Singh, V. K.; Doreswamy, R. and Saini, M. (2010). Molecular Characterization of Heat Shock Protein 70-1 Gene of Goat (Capra hircus). Mol Biol Int. 108429.
  6. Hall, D.M.; Xu, L.; Drake, V.J.; Oberley, L.W.; Oberley, T.D.; Moseley, P.L. and Kregel, K.C. (2000). Aging reduces adaptive capacity and stress protein expression in the liver after heat stress. J Appl Physiol. 89:749-759.
  7. Legare, C.; Thabet, M. and Sullivan, R. (2004). Expression of heat shock protein 70 in normal and cryptorchid human excurrent duct. Mol Hum Reprod. 10:197-202.
  8. Liu, Y.X.; Li, D.Q.; Cui, Q.W.; Shi, H.X. and Wang, G.L. (2010). Analysis of HSP70 mRNA level and association between linked microsatellaite loci and heat tolerance traits in dairy cows. Yi Chuan. 32:935-941.
  9. Mishra, A.; Hooda, O.K.; Singh, G. and Meur, S.K. (2010). Influence of induced heat stress on HSP70 in buffalo lymphocytes. J Anim Physiol Anim Nutr (Berl). 22:1111-13.
  10. Morimoto, R. and Fodor, E. (1984). Cell-specific expression of heat shock proteins in chicken reticulocytes and lymphocytes. J cell boil. 4:1316-23.
  11. Patir, H. and Upadhyay, R.C. (2007). Interrelationship between Heat Shock Protein 70 (HSP70) and Lymphocyte Proliferation in Thermal Exposed Buffalo Heifers. Ita J of Anim Sci. 6:1344-1346.
  12. Patir, H. and Upadhyay, R.C. (2010). Purification, characterization and expression kinetics of heat shock protein 70 from Bubalus bubalis. Res Vet Sci. 88:258–262.
  13. Wang, S.; Xie, W.; Rylander, M.N.; Tucker, P.W.; Aggarwal, S. and Diller, K.R. (2008). HSP70 kinetics study by continuous observation of HSP–GFP fusion protein expression on a perfusion heating stage. Biotechnol. Bioeng. 99:146–154.
  14. Zulkifli, I.; Norbayyah, B.; Cheah, Y.W.; Soleimani, A.F.; Sazili, A.Q.; Goh, Y.M. and Rajion M.A. (2010). A note on heat Shock protein 70 expression in goats subjected to road transportation under hot, humid tropical conditions. Animal. 4:973-976.

Global Footprints