Electroporated sperm mediated gene transfer in Indian major carps
N. Sarangi1, A.B. Mandal1*, A.K. Bandyopadhyay1, T. Venugopal2, S. Mathavan2 and T.J. Pandian2
1Biotechnology Laboratory, Central Agricultural Research Institute, Port Blair 744 101, India.
2 School of Biological Sciences, Madurai Kamaraj University, Madurai, Tamil Nadu 625 014, India.
(Received 10 April 1999 / Accepted 19 October 1999)
Abstract.
The rainbow trout growth hormone (rtGH) gene under the transcription control of long terminal repeat of Rous sarcoma virus was successfully introduced into three Indian major carps (IMC) viz. rohu (Labeo rohita) catla (Catla catla) and mrigal (Cirrihinus mrigala), en route electroporated sperms. The present communication enumerates the results of an array of electroporation experiments aimed at standardising diverse variables like voltage, capacitance, resistance and pulse constant to achieve maximum transformation efficiency. Stable genomic integration of the intruded alien gene was demonstrated through slot blot hybridization in all the three species. Per cent transgenic individuals were largely varied in the different species, maximum of 25% being observed in rohu followed by 23 and 13% in mrigal and catla, respectively. This indicates that electroporated sperm mediated gene transfer may emerge as a convenient means in fish transgenic development specially in IMC. Optimisation of electroporation condition for rtGH transformation to produce transgenic IMC are discussed hereunder in detail.
Keywords: rtGH, electroporation, sperm mediation, Indian major carps
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