Transgene expression in Penaeus monodon cells: evaluation of recombinant baculoviral vectors with shrimp specific hybrid promoters
- 274 Downloads
It has been realized that shrimp cell immortalization may not be accomplished without in vitro transformation by expressing immortalizing gene in cells. In this process, efficiency of transgene expression is confined to the ability of vectors to transmit gene of interests to the genome. Over the years, unavailability of such vectors has been hampering application of such a strategy in shrimp cells. We report the use of recombinant baculovirus mediated transduction using hybrid promoter system for transgene expression in lymphoid cells of Penaeus monodon. Two recombinant baculovirus vectors with shrimp viral promoters (WSSV-Ie1 and IHHNV-P2) were constructed (BacIe1-GFP and BacP2-GFP) and green fluorescent protein (GFP) used as the transgene. The GFP expression in cells under the control of hybrid promoters, PH-Ie1 or PH-P2, were analyzed and confirmed in shrimp cells. The results indicate that the recombinant baculovirus with shrimp specific viral promoters (hybrid) can be employed for delivery of foreign genes to shrimp cells for in vitro transformation.
KeywordsTransgene expression Penaeus monodon Shrimp specific vectors Hybrid promoter Baculoviral vectors
This research was supported by the Department of Biotechnology, Government of India (BT/PR8050/AAQ/03/289/2006 and BT/PR5126/AAQ/3/591/2012). The first author thanks DBT for Fellowship.
- Carbonell LF, Klowden MJ, Miller LK (1985) Baculovirus-mediated expression of bacterial genes in dipteran and mammalian cells. J Virol 56:153–160Google Scholar
- Felder JM, Sutton RE (2009) Lentiviral vectors. In: Templeton NS (ed) Gene and cell therapy. CRC Press, Boca Raton, pp 207–243Google Scholar
- Guo R, Yifan Z, Sheng L, Haoping X, Miao Z, Biao L (2010) Sodium butyrate enhances the expression of baculovirus-mediated sodium/iodide symporter gene in A549 lung adenocarcinoma cells. Nucl Med Commun 31:916–921Google Scholar
- Huard J, Lochmuller H, Acsadi G, Jani A, Massie B, Karpati G (1995) The route of administration is a major determinant of the transduction efficiency of rat tissues by adenoviral recombinants. Gene Ther 2:107–115Google Scholar
- Jayesh P (2013) Development of lymphoid cell culture system from Penaeus monodon and molecular approaches for its transformation. Ph.D. Thesis. Cochin University of Science and Technology, IndiaGoogle Scholar
- Jayesh P, Philip R, Singh ISB (2015) Multifactorial interaction of growth factors on Penaeus monodon lymphoid cells and the impact of IGFs in DNA synthesis and metabolic activity in vitro. Cytotechnology 67:559–571Google Scholar
- Lesch HP, Laitinen A, Peixoto C, Vicente T, Makkonen K-E, Laitinen L, Pikkarainen JT, Samaranayake H, Alves PM, Carrondo MJT, Ylä-Herttuala S, Airenne KJ (2011) Production and purification of lentiviral vectors generated in 293T suspension cells with baculoviral vectors. Gene Ther 18:531–538CrossRefGoogle Scholar
- Luo W-Y, Shih Y-S, Lo W-H, Chen H-R, Wang S-C, Wang C-H, Chien C-H, Chiang C-S, Chuang Y-J, Hu Y-C (2011) Baculovirus vectors for antiangiogenesis-based cancer gene therapy. Cancer Gene Ther 18:637–645Google Scholar
- O’Reilly DR, Miller LK, Luckow VA (1992) Baculovirus expression vectors. A laboratory manual. Freeman WH, New YorkGoogle Scholar
- Smith GE, Summers MD (1989) Method for producing a recombinant baculovirus expression vector. US Patent, 4879236Google Scholar