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Abstract This paper discovered the growth improvement of red carp transferd with the myostatin propeptide gene from Larimichthys crocea . Myostatin (MSTN) propeptide gene from Larimichthys crocea and the internal ribosome entrysite(IRES)-enhanced green fluorescence protein gene(EGFP) fragment from the pIRES-EGFP plasmid were cloned by RT-PCR and PCR, respectively. After confirming their sequences, the recombinant Tol2 transposon donor plasmid pT2AL200R150G-MSTN propeptide-IRES-EGFP was constructed. Three different gene transfer methods, sperm-mediated gene transfer (SMGT), electroporation (E) and gene gun (G) were used to obtain transgenic red carp (Cyprinus carpio). In SMGT, common sperm storage buffer (S1) and hypotonic dilution (S2) were used, respectively. In electroporation, 1 500 V (E1) and 2 500 V (E2) were applied, respectively. The rates of EGFP expression were as follows: S1, 38%; S2, 48%; E1, 47%; E2, 53% and G, 2% in larvae after 72 of hatching, respectively. The positive rates of reverse transcript-polymerase chain reaction (RT-PCR) for the transgene were as follows: S1, 35%; S2, 45%; E1, 45%; E2, 55% and G, 1.8% in larvae after 10 d of hatching, respectively. After 75 d of hatching, the body length and weight of transgenic fish increased 21.31% and 27.59% in comparison with the control, respectively. The results showed that gene transfer efficiency could be improved by treating sperm with hypertonic and hypotonic dilution, and electroporation.
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Received: 12 July 2012
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