• Journal of Sericultural and Entomological Science
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• v.33 no.1
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• pp.1-5
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• 1991

PBS soluble- and insoluble-extracts form silkworm eggs were analysed by native-PAGE and SDS-PAGE. During the embroyonic development, soluble proteins which are mainly composed of ESP, vitellin and 30K proteins showed similar degradation pattern in both electrophoretic analyses. Several peptides which seemed to be intermediated forms of yolk proteins were detected in latter part of embryogenesis, while a protein band newly appeared in one day elapsed after acid treatment. When insoluble extracts from silkworm eggs were analysed with SDS-PAGE, several peptides were detected at the later stages of the embryonic development, and newly hatched larvae. These peptides are considered to be structure proteins for embryonic morphogenesis.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2000.10a
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• pp.68-68
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• 2000

• Journal of Sericultural and Entomological Science
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• v.51 no.1
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• pp.73-77
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• 2013

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.

• Korean journal of applied entomology
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• v.61 no.1
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• pp.249-254
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• 2022

Silkworms, which have for long been used as an insect resource for industrialization, have recently attracted attention as potential bio-factories for the production of novel biomaterials. In this regard, material production is typically achieved based on transformation technology, mediated via microinjection, in which a target gene is inserted into eggs containing an embryo. However, an essential step in the microinjection procedure is egg fixation, which can be a time-consuming and laborious task. Therefore, in this study, using the 3DCADian program, we adopted a 3D printing approach to model egg liners and glue drawers, which can contribute to facilitating egg alignment and fixation, thereby enhancing transformation efficiency by reducing time consumption and fatigue. After rendering using Fusion 360, the two supplementary tools were produced by printing with nylon resin (PA12) and Sinterit Lisa Pro. Subsequent analysis of the time required to fix eggs on glass slides using the two manufactured tools, revealed that the processing time was reduced by approximately 18.6% when the two tools were used compared with when these tools were not used. These innovations not only reduced fatigue but also contributed to more effective use of the microscope and manipulator for microinjection. Consequently, we believe that with additional research and refinement, the egg liner and glue drawer developed in this study could be used to enhance silkworm transformation efficiency and study similar transformation systems in other industrial insects.

• Korean journal of applied entomology
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• v.53 no.3
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• pp.199-207
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• 2014

It is true that the proper environmental risk assessments for many GM (Genetically Modified) insects almost have not been executed in Korea. Therefore, we tested the environmental risk assessment about GM silkworms if there is any difference between GM silkworms and non-GM silkworms by the following three measurements. First, we measured their mobility in the breeding environment conditions with food and without food. Secondly, we measured their viability at the artificial extreme environmental conditions (low and high temperature and humidity, absent/present of foods,) after escaping from their breeding environments. Thirdly, we observed the number of laying eggs and their hatchability between GM silkworms and non-GM silkworms with four different pair experiments. The mobility of GM silkworms and non-GM silkworms statistically did not differ, and the egg productivity and hatchability were not also different. The hatchability by couple of GM female silkworms and non-GM male silkworms was lower than by non-GM male and female couple between the GM silkworms and non-GM silkworms, and there was statistically different. Relatively, the viability of GM silkworms was lower than non-GM silkworms. We could not exactly test for viability of silkworms in low temperature conditions because of their hibernating. Although there was any difference in viability and hatchability between GM silkworms and non-GM silkworms, all ability of GM silkworms was lower than non-GM silkworms. Conclusively, the environmental risk of GM silkworm was relatively lower than non-GM silkworm in this study.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.151-154
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• 2000

The effect of silkworm hemolymph on insect cell apoptosis was investigated. The addition of silkworm hemolymph into the culture medium either before or during the baculovirus infection increased the host cell longevity. Silkworm hemolymph also inhibits apoptosis induced by actinomycin D, the RNA synthesis inhibitor. In this study, a flow cytometry was used for the quantitative analysis of apoptosis inhibition by silkworm hemolymph.

• Journal of Sericultural and Entomological Science
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• v.53 no.1
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• pp.55-60
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• 2015

Melittin is the main component of Bee Venom and has antibacterial activity against several bacteria. To produce the melittin antimicrobial peptide, we constructed transgenic silkworm that expressed melittin gene under the control BmA3 promoter using piggyBac vector. The use of the 3xP3-driven EGFP cDNA as a marker allowed us to rapidly distinguish transgenic silkworm. Mixtures of the donor vector and helper vector were micro-injected into 300 eggs of bivoltin silkworms, Baegokjam. In total, 131 larvae (G0) were hatched and allowed to develop into moths. The resulting G1 generation consisted of 36 broods, and we selected 4 broods containing at least 1 EGFP-positive embryo. The rate of successful transgenesis for the G1 broods was 11%. We identified 12 EGFP-positive G1 moths and these were backcrossed with wild-type moths. With the aim of identifying a melittin as antimicrobial peptide, we investigated the Radical diffusion Assay (RDA) and then demonstrated that melittin possesses high antibacterial activities against gramnegative bacteria.

• Journal of Sericultural and Entomological Science
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• v.51 no.2
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• pp.207-210
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• 2013

In this study, matured silkworm hemolymph was studied for inhibitory effect on melanin formation, tyrosinase activities. A matured silkworm hemolymph mass purification method was developed and some of suppressing melanin formation effects were investigated, too. As a result, the advanced purification method was higher in yield rate than existing one by 53%. A free radical scavenging activity of hemolymph was higher than sericin on the market($IC_{50}$, $202.9{\pm}33.1$) and lower than vitamin C($0.57{\pm}0.02$) by $6.4{\pm}0.3{\mu}g/m{\ell}$. An inhibitory effect on melanin synthesis was higher in hemolymph compared to Morus bark and arbutin by 9.15% and 11.56%, respectively. An inhibitory effect on tyrosinase activity was higher in hemolymph than Morus bark and lower than arbutin. Effect of hemolymph $45{\mu}g/m{\ell}$ on suppressing of tyrosinase was almost 90%. Thus, these results suggest that hemolymph of matured silkworm may have beneficial properties as a material for cosmetics.

• Journal of Life Science
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• v.21 no.12
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• pp.1726-1731
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• 2011

Human Stem Cell Factor (hSCF) is a cytokine that binds to the c-Kit receptor and plays an important role in hematopoiesis, spermatogenesis, and melanogenesis. To produce the human Stem Cell Factor (hSCF) recombinant protein, we constructed a germline transgenic silkworm using the piggyback vector. The expression of the hSCF gene was driven by the Drosophila heat shock protein 70 (dHsp70) promoter. 3XP3 promotor-driven EGFP was used as a marker which allowed us to rapidly distinguish the transgenic silkworm. A mixture of the donor and helper vector was micro-injected into 1,020 eggs of bivoltin silkworms, Keomokjam. We obtained approximately 22 G1 broods that were EGFP-positive. The expression of the hSCF gene in the transgenic silkworm was analyzed by SDS-PAGE and immunoblotting. Also, analysis of insertion sites into the silkworm genome using inverse PCR showed that exogenous DNA was inserted into the transgenic silkworm genome. These results show that successfully constructed transgenic silkworm expresses the hSCF recombinant protein.

• Korean journal of applied entomology
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• v.41 no.2
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• pp.85-89
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• 2002

Development and reproduction of the rice armyworm, Pseudaletia separara, were investigated on different diets (corn leaf, rice leaf, Chinese cabbage leaf, cabbage leaf, sweetpotato leaf, soybean leaf and silkworm artificial diet). Egg and pupal periods were not different among the diets. But larval periods showed significant difference among diets. Developmental period from egg to adult was shortest as 32.5 d on corn leaf and longest as 46.6 d on soybean leaf. At 16 days after hatching, larval weights on rice and silkworm artificial diet were heavier than those on other diets. The larvae reared on cabbage leaf and sweetpotato leaf failed to pupate. Survival rates from larva to adult were highest (70.6%) on com leaf lowest (39.8%) on soybean leaf. The longevity of female adults was not affected by the larval diets. Female adults showed higher fecundity when the larvae were fed with corn leaf, rice leaf and silkworm artificial diet than other diets. Net reproduction rate (R$_{0}$) was highest as 1218.5 on silkworm artificial diet. The intrinsic rate of natural increase (R$_{0}$) ranged from 0.115 to 0.175, and was maximum on com leaf.