| Summary: | 碩士 === 國立中興大學 === 昆蟲學系所 === 99 === In this study, the transposon, piggyBac, was used as the transgenic vector, to transform the oriental fruit fly Bactrocera dorsalis (Hendel). The target protein, recombinant chicken growth hormone (cGH), is under the control of Drosophila melanogaster heat shock protein 70 (hsp70) promoter. Total of 1,861 embryos were microinjected, and finally one G0 male was obtained with DsRed (reporter gene) expressing in the G1 progeny, resulting a transformation efficiency of 4 %. The piggyBac-mediated integration was confirmed by Southern blotting and inverse PCR analysis. At least four copies of transgenic expression cassettes were integrated into the transformed B. dorsalis genome, and the results of inverse PCR further revealed that three of four expression cassettes were integrated on the non-coding sequences. Moreover, the cGH expression was further detected by RT-PCR and Western blotting analysis. The results of RT-PCR revealed the tight regulatory of hsp70 promoter expression. The RNA expression of cGH as detected at 2-, 12-, 24-, 36-, and 48-h after heat shock induction at both 35 and 37oC, while the protein was only detected at 48 h. The differences in expression profiles between protein and mRNA might be cause by heat shock response that blocks Cap-polyA dependent translation.
|
|---|
