摘要
Ecological safety is a major consideration in the commercialization of transgenic fish. Development of sterile transgenic triploid fish through hybridization of transgenic tetraploid fish and transgenic diploid fish is a feasible way to solve this problem. The "all-fish" transgene, pbcAbcGHc, containing the black carp β-actin gene promoter and the open reading frame (ORF) of the black carp growth hormone (GH) gene was constructed and introduced into fertilized eggs of allotetraploid fish through microinjection. Contrast cultivation results showed that the growth rate of 150 day-old P 0 black carp GH gene transgenic allotetraploid fish was much higher than that of controls. Sixty 150 day-old transgenic allotetraploid fish were assayed by PCR for transgene integration and 90% of fish were positive for the transgene. The transgene was detected in 13 of 20 sperm samples from male transgenic allotetraploid fish. RT-PCR detected transcription of the exogenous black carp GH gene in the muscle, liver, kidney and ovaries of the largest transgenic allotetraploid fish. This study has developed P 0 black carp GH gene transgenic allotetraploid fish with a highly increased growth rate, which provides a solid foundation for the establishment of a pure line of transgenic allotetraploid fish and for the large scale production of sterile transgenic triploid fish.
Ecological safety is a major consideration in the commercialization of transgenic fish. Development of sterile transgenic triploid fish through hybridization of transgenic tetraploid fish and transgenic diploid fish is a feasible way to solve this problem. The ＂all-fish＂ transgene, pbcAbcGHc, containing the black carp β-actin gene promoter and the open reading frame （ORF/of the black carp growth hormone （GH） gene was constructed and introduced into fertilized eggs of allotetraploid fish through microinjection. Contrast cultivation results showed that the growth rate of 150 day-old P0 black carp GH gene transgenic allotetraploid fish was much higher than that of controls. Sixty 150 day-old transgenic allotetraploid fish were assayed by PCR for transgene integration and 90% of fish were positive for the transgene. The transgene was detected in 13 of 20 sperm samples from male transgenic allotetraploid fish. RT-PCR detected transcription of the exogenous black carp GH gene in the mus cle, liver, kidney and ovaries of the largest transgenic allotetraploid fish. This study has developed P0 black carp GH gene transgenic allotetraploid fish with a highly increased growth rate, which provides a solid foundation for the establishment of a pure line of transgenic allotetraploid fish and for the large scale production of sterile transgenic triploid fish.
基金
supported by the National Special Fund for Scientific Research in Public Benefits (Grant No. 20090304608)
the National Basic Research Program of China (Grant No. 2007CB109006)
the National Natural Science Foundation of China (Grant No. 30500294)
the Excellent Talent Program of Hunan Normal University (Grant No. ET31004)
