Transgenesis of silkworms has great potential for the development of silk with new properties as well as for the preparation of recombinant proteins for the use in biomedicine. Zabelina et al. (2015a) showed that transgenesis of parthenogenetic silkworms facilitates the selection and maintenance of transgenic homozygotes with stabile transgene insertions. However, the efficiency of transgenesis was less than 2 % compared to 60 % in the standard, non-parthenogenetic silkworms. The purpose of the present research was to explore the cause of this difference. Since transgenesis is normally performed at 25 °C but in the parthenogenetic silkworms at 15 °C (3 days incubation at this temperature is part of the protocol for the induction of parthenogenetic development), we assumed that the eggs incubated at 15 °C might have been injected with the DNA construct at unsuitable time. The work was therefore focused on the rate of embryogenesis at 15 °C in the eggs treated in different ways. Intensive cleavage of the control eggs (strain K23) was observed at 12 h after oviposition at 25 °C and between 24 and 36 h at 15 °C. The transgenesis of parthenogenetic silkworms is also complicated by the embryonic diapause. In the current work, diapause was suppressed by implanting PK1 ovaries into the non-parthenogenetic male hosts K23. Parthenogenetic development was activated by the heat shock in the chorionated eggs dissected from the implants. No cleavage was detected at 12 h after the acti-vation and nearly complete blastoderm was found at 48 h. In respect to the course of cleavage at 15 °C, transgene injection 24 h after the activating heat shock can be recommended. The eggs from endogenous ovaries of the K23 females, which also contained the implants of the PK1 ovaries, acquired partial capacity of parthenogenesis. Low rate of embryogenesis was also found in the transgenic clone VTG1. Current results suggest that more research is needed to understand and possibly explore differences in the rate of silkworms exposed to different treatments.
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:320624 |
| Date | January 2017 |
| Creators | VRCHOTOVÁ, Markéta |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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