Modification of the commercial silkworm eggs adequate for Bluemoon0silkworm transgenesis

Silkworm transgenesis scientists have done some genetic modification work on multivoltine silkworms, but that type of silkworms is less commercial feasible. They are easy to manipulate, because they breed all year round. But the commercial silkworm variety must undergo hydrochloric acid treatment at a high temperature to be artificially hatched. Hydrochloric acid penetrates through the holes in the silkworm eggs, fatally damaging their reproduction. So it had been thought that altering the properties of the commercial silkworm variety would be very difficult. So we have tried to make from diapause to non-diapause eggs using diapauses varieties, 'Backokjam' and 'Jam 124'. At present, our group has establishing the conditions for non-diapause eggs. Oviposited eggs after 40 ~ 60 hours were incubated for 24 hours at $15{\sim}20^{\circ}C$ with dark condition. Non-diapause eggs were completely induced. The hatching rate, molting rate and pupation rate of non-diapause 'Jam 124' and 'Backokjam' eggs showed no differences compared to diapause eggs. When transgenic silkworm using the non-diapause eggs, the hatching rate showed that non-diapause eggs induced from diapause were 40 ~ 70%, diapause eggs treated with artificial incubation were 10 ~ 30%, and polyvoltine strains, HM eggs were 30 ~ 50%. Therefore, we suggest that modification techniques of the commercial silkworm eggs adequate for silkworm transgenesis can be used to develop transgenic silkworms more easily.

누에 형질전환에 적합한 실용품종 누에알의 제조

지금까지 형질전환에 사용된 품종은 휴면각성을 위해 염산처리가 불필요한 비휴면계통인 다화성품종에서 제한적으로 이뤄지고 있으며, 휴면계통의 누에알을 대상으로 형질전환누에를 제작한다는 것은 매우 어려운 것으로 판단되었다. 따라서, 본 연구에서는 양잠농가에 장려품종으로 보급되고 있는 백옥잠(잠123 ${\times}$ 잠124)과 원종인 잠124의 월년란을 불월년란화 하기 위한 처리조건을 구명한 바 산란 후 40 ~ 60시간째에 온도는 $15{\sim}20^{\circ}C$, 조명은 암(0L:24D)처리로 100% 불월년화가 유도되었다. 또한 불월년란은 월년란에 비해 부화율, 상족율 및 화용비율에 있어 큰 차이가 없었다. 불월년란으로 유도된 실용품종 누에알에 누에 형질전환용 전이벡터를 미세주입한 후 부화율을 분석한 결과, 불월년란으로 유도된 월년란의 부화율은 40 ~ 70% 로, 인공부화 처리한 월년란의 10 ~ 30%, 다화성누에 HM의 30 ~ 50% 부화율에 비해 높은 부화율을 나타내었다. 본 연구를 통해 개발한 불월년란 유도 기술은 현재 양잠 농가에 보급하고 있는 실용품종을 형질전환하는데 매우 유용하게 사용될 것으로 기대된다.