2010 Annual Report
Using the technology of androgenesis, the following homozygous clonal lines of transgenic fish containing cecropin P1 constructs were created: S7-342-F695, S8-505-G231, S7-375-F180, S9746-F509 and S9-659-F-073 which are resistant to IHNV; and A. salmonicida, and S9-638-F297, U6-768-G410, A12-944 and A13-831 which are resistant to IHNV alone. Through genotyping and sex identification by PCR analysis, all male individual homozygous fish have been tagged. Individuals from families S7#375-F-073 and S9#659-F-180 have reached reproductive maturation. The other families will reach reproductive maturation by December 2010. The expression of cecropin P1 transgene in heart, spleen, muscle and liver of individual fish from S7#375-F-073 and S9#659-F-180 have been confirmed by RT-PCR analysis.
Sperm samples from S7#375-F-073 and S9#659-F-180 homozygous transgenic fish families have been cryopreserved.
Sperm of S7#375-F-073 and S9#659-F-180 were out-crossed to eggs from non-transgenic fish and the resulting offspring were challenged with Aeromonas salmonicioda. Results of the challenge studies showed that progeny derived from both families of fish exhibited resistant characteristics to Aeromonas salmonicidas.
A construct which contains crtW and crtZ genes driven by the Beta-actin gene promoter has been constructed. This double transgene construct is transfected into CHSE cells (Chinook salmon embryonic cells) to test for conversion of Beta-carotene into astaxanthin by HPLC analysis. The transgene has been proven to be functional. The transgene insert will be introduced into rainbow trout via electroporating the sperm following conditions developed in our laboratory. The resulting fish will be raised to adulthood for identification of the presence of transgene by PCR analysis.
The ADODR is in frequent contact with the cooperator through phone calls, email, and site visits in addition to receipt of written reports.