Production of Transgenic Homozygous Diploid in Mud Loach(Misgurnus mizolepis) I. Transfer of Luciferase Gene and Evaluation of Mud Loack Expression Vector

  • Nam Yoon Kwon (Department of Aquaculture and Pukyong National University) ;
  • Kim Moo-Sang (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Lee Hyung-Ho (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Kim Dong Soo (Department of Aquaculture and Pukyong National University)
  • Published : 1996.08.01

Abstract

Validities of several gene transfer methods including microinjection, electroporation and lipo-fection with luciferase gene (pRSVL), and effectiveness of mud loach expression vector which contains ARS from mud loach on production of transgenic mud loach were evaluated. Microiniection revealed the $0\~8\%$ of transgene incidence in 2-week-old fish with significant mosaicism. Electroporation and lipofection of mud loach sperm also successfully introduced the transgene into sperm cells, and transferred the foreign DNA into zygote. Gene transfer by electroporation and lipofection showed a range of $0\~28\%$ and $0\~48.1\%$ of transgene incidence, respectively in newly hatched larvae, altough most DNA introduced were gradually degraded with the development of fish. Microinjections of mud loach expression vector caused a significantly reduced survival rate of mud loach embryos with severe teratogenic effects, and ARS/Luc transgene could not be detected in normally developed fish after microinjection.

Keywords