• Title/Summary/Keyword: Cene transfer by electroporated sperm

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Transgenesis in Fish: Indian Endeavour and Achievement

  • Pandian, T.J
    • Journal of Aquaculture
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    • v.16 no.1
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    • pp.51-58
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    • 2003
  • The first Indian transgenic fish was generated in 1991 using borrowed constructs from foreign sources. To construct transformation vectors for the indigenous fishes, growth hormone genes of rohu (r-CH), Labeo rohita and catfish, Heteropneustes fossilis were isolated, cloned and sequenced; their fidelity was confirmed in prokaryotic and eukaryotic systems. A vector was constructed with grass carp b-actin promoter driving the expression of r-GH. Rohu eggs are large. fragile and swell 2~3 times. when fertilized. Hence they were amenable only for electroporated sperm-mediated gene transfer. Accordingly, the sperm electroporation technique was standardized to ensure 25% hatchling survival and 37% Presumptive transgenics without suffering any deformity. Southern analysis confirmed genomic integration in 15% of the tested individuals (Ti) belonging to family lines 2 and 3: another 25% of the Juveniles (Te) were also proved transgenic but with the transgene persisting extrachromosomally for longer than 1 to 2 years. perhaps due to the presence of replicon in the vector. Transgenics belonging to different family lines grew 6~8 times faster than the respective controls. Difference in growth trends of Ti and Te within a family line was not significant. In the Ti family 3 remarkable growth acceleration was sustained for a period longer than 36 weeks but in those of family 2, it gradually decreased. All transgenic fishes including the rohu converted the food at a significantly higher efficiency. Barring the transgenic mudloach, all the other transgenic fishes consumed food at significantly reduced rate.