• International Journal of Industrial Entomology and Biomaterials
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• 제7권2호
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• pp.221-229
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• 2003

Ten multivoltine female parental lines were crossed with popular bivoltine male silkworm breed NB$_4$D$_2$. Three types of heterosis parameters viz., heterosis over mid-parental value (hybrid vigour), heterobeltiosis (useful heterosis) and standard heterosis (standard check) were estimated for 15 economically important quantitative traits. The interaction among the hybrids and parents indicated significant effect for maximum characters. The heterotic effect of new hybrid combination was compared with popular hybrids viz., Pure Mysore${\times}$NB$_4$D$_2$ and Nistari${\times}$NB$_4$D$_2$. Varied heterotic effect was observed for different traits for different hybrid combination. The results inferred that the crosses viz., BL$_{23}$${\times}$NB$_4$D$_2$ ranked top for 14 traits followed by Hosa Mysore${\times}$NB$_4$D$_2$ for 11 traits; PA$_{12}$${\times}$NB$_4$D$_2$ for 9 traits; BL$_{24}$${\times}$NB$_4$D$_2$ for 8 traits; Kolar Gold${\times}$NB$_4$D$_2$ for 7 traits; WAI$_1$${\times}$NB$_4$D$_2$ for 6 traits and MU$_{11}$${\times}$NB$_4$D$_2$ for 5 traits. Among these, the best hybrids Kolar Gold${\times}$NB$_4$D$_2$ and MU$_{11}$${\times}$NB$_4$D$_2$ were identified for longer filament length and fine denier. Similarly for higher cocoon yield and silk productivity BL$_{223}$${\times}$NB$_4$D$_2$ and BL$_{24}$${\times}$NB$_4$D$_2$ were found to be superior. These hybrid combinations are suitable for commercial exploitation at large scale.e.e.e.

• International Journal of Industrial Entomology and Biomaterials
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• 제2권2호
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• pp.123-127
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• 2001

In order to introduce bivoltine races in a tropical country like India, it is necessary to have stability in cocoon crop under high temperature environments. Unlike any temperate country like Japan, the rearing conditions/environment, climatic conditions, quality of mulberry leaf and incidence of diseases are unpredictable in India. Geneticists and breeders of all the sericultural countries have experienced the influence of environment during the process of breeding. in order to select efficiently the breeds with high temperature tolerance, it is verb important to analyse clearly the heritability nature of high temperature tolerance. In light of the above, the present study was undertaken to determine the effect of high temperature treatment of (A) $35{\pm}1^{\circ}C$ and 85${\pm}$5% RH for 24 hrs continuously, (B) $35{\pm}1^{\circ}C$ and 85${\pm}$5% RH for 48 hrs continuously and (C) the control (25${\pm}1^{\circ}$ and 65${\pm}$5% RH in the normal rearing condition from the $3^{rd}of5^{th}$ instar on the pure races such as Moria, Nl37 and Cl46 as well as their Fl hybrids. The overall performance indicate that the hybrids are mare tolerant than the pure races. it was also observed that the overall performance declined in those batches where 48 hrs treatment was given. The most interesting observation noticed in this study was that there was maternal effect regarding temperature tolerance as evident from the better performance of those hybrids where the female parent used was more tolerant as pure race.

• International Journal of Industrial Entomology and Biomaterials
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• 제2권2호
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• pp.107-110
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• 2001

Variations in protein and nucleic acid concentrations were observed in 24 hrs old eggs and hatched larvae of Nistari strain, Bombyx mori, exposed to starvation. Three starvation treatments of 24,48 and 60 hrs were given separately from 0 hr old fifth instar larvae. Biochemical variations were studied in the resultant hatched larvae of one time starved parent, while the eggs obtained from parents receiving starvation in two successive generations were considered for the study. In hatched larvae, protein levers in 24 hrs starvation groups remained significantly higher over control (never starved) while the same was found to be lower in 48 and 60 hrs starvation individuals. The RNA concentration remained significantly higher in all the treated lots. However, DNA content was not found to be significantly altered in hatched larvae after exposure to feeding stress. Protein, RNA and DNA concentration of 24 hrs old eggs produced by all the starved groups of Nistari, which had deceived two consecutive starvation during parental generations, showed higher concentrations of these biomolecules over control. Hence, starvation induced alterations in protein and nucleic acids in eggs and hatched Iarvae are indicative of a preparatory phase adopted by the insect to acclimatise itself and its progeny to stress situations.

• 한국잠사곤충학회지
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• 제40권2호
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• pp.111-116
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• 1998

To construct transformed Bm5 cells, Autographa californica nuclear polyhedrosis virus (AcNPV)IE1 gene, an immediate early viral gene was firstly used in this study. AcNPV IE1 gene, which shares on 95.3% uncleotide sequence homology with Bombyx mori nuclear polyhedrosis virus (BmNPV) IE1 gene, was isolated and cloned into pBluescript. Neomycin gene from pKO-neo was inserted under the control of the IE1 promoter to yield pAcIE1-neo. The plasmid pAcIE1-neo was transfected into Bm5 or Sf9 cells, and neomycin-resistant cells were selected in TC100 medium containing 10% fetal bovine serum (FBS) and 1 mg/$m\ell$ G418 for two weeks. Individual clones were picked and each was amplified for further characterization. The genomic DNA from neomycin-resistnt cells was isolated and characterized by PCR using AcNPV IE1 gene-specific primers and by Southern blot analysis using neomycin gene probe. We concluded that AcNPV IE1 gene was functional in B. moridrived Bm5 cells as well as Spodaptera frugiperda-derived Sf9 cells to produce stably-transformed insect cells.

• 한국잠사곤충학회지
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• 제46권1호
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• pp.28-31
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• 2004

견피브로인의 응용가능성을 높이기 위하여 대표적인 생체고분자인 히아론산을 이용하여 견피브로인/히아론산 브렌드 필름을 제조하였으며 구조 및 열 특성을 살펴보았다. 1. 적외선 분광분석 결과 견피브로인과 히아론산 두 고분자 사이에는 수소결합 등의 상호작용이 존재하지 않는 것으로 나타났다. 또한 브렌드 필름내에 존재하는 실크피브로인은 EDC 에탄올 용액처리에 의하여 $\beta$-sheet 구조로 전이되었다. 2. 견피브로인/히아론산 브렌드 필름에서 견피브로인과 히아론산 각각의 열분해 흡열피크와 발열피크 온도에는 큰 변화가 없었으나, 열분해 피크의 폭이 넓어지는 것으로 봐서 실크 피브로인과의 브렌드 또는 EDC 처리에 의하여 열분해 반응이 매우 복잡한 환경하에서 진행되고 있음을 알 수 있었다. 3 견피브로인/히아로산 브렌드 필름은 자기 중량 대비 평균 70배의 수분을 흡수하였다.

• International Journal of Industrial Entomology and Biomaterials
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• 제27권2호
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• pp.282-288
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• 2013

In nature, the population of Nosema bombycis (Microsporidia) causing pebrine disease is small and their development is extremely slow and only few ultimately producing spores. Pebrine infected silkworm, Bombyx mori larvae collected from sericulture field were alive till $3^{rd}$ generation though the concentration of N.bombycis spore was very high ($2.4-3.0{\times}10^8$ spores. $mL^{-1}$). All larvae were died during $4^{th}$ generation with extremely high concentration of pebrine spores ($3.0-4.0{\times}10^9$ spores. $mL^{-1}$) and mostly contain long polar tube (LT). Alternately, all larvae were died immediately (at $3^{rd}$ stage of $1^{st}$ generation) when it was artificially inoculated with same concentration of N.bombycis spores harvested from field ($2.4-3.0{\times}10^8$ spores. $mL^{-1}$) though concentration of spores harvest was very less ($3.0-4.0{\times}10^6$ spores. $mL^{-1}$) and mostly contain short polar tube (ST). Artificially pebrine infected male moth when mated with healthy female moth took six generations to develop pebrine disease and all larvae were died at the $2^{nd}$ stage with very less spore harvest ($3.0-10.0{\times}10^6$ spores. $mL^{-1}$). Survival percentage was increased in all generations (~92.0% at $4^{th}$ generation) when silkworm rearing was conducted under new integrated disease management system.

• 생명과학회지
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• 제13권5호
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• pp.596-603
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• 2003

누에배양세포주(Bm5)에 N-glycosylation 저해제인 tunicamycin를 처리하여 인위적으로 UPR를 유도하고 이로부터 cDNA 유전자은행을 제작한 후, 정상세포주에 비하여 발현량이 증가하는 40개의 차별화 발현 cDNA 클론을 선발하였다. 차별화발현 클론 중에서 기존에 밝혀진 전사제어인자와 1차 아미노산의 구조적 유사성을 나타내는 클론(ATFC)에 대하여 유전자의 구조와 발현특성을 분석한 결과는 다음과 같다. 유전자의 구조분석 결과, ATFC는 효모의 전사제어인자 Hac1p와 구조적으로 매우 유사하게 $\alpha$-helix 상의 7개 아미노산 잔기 마다 leucine이 7회 반복하여 출현하는 leucine zipper 모티프가 존재하고 있었으며, leucine zipper 바로 앞쪽에는 분자 샤페론이나 folding enzyme의 프로모터 부위에 존재하는 UPRE에 결합할 것으로 추정되는 염기성 아미노산이 풍부한 basic region이 존재하고 있었다. 또한, ATFC 유전자에 대하여 분자샤페론 및 folding enzyme의 전사 촉진 기능을 해석하기 위하여 누에 배양세포주(Bm5)에 각종 스트레스 유도제를 처리한 후 ATFC의 발현특성을 분석한 결과, 정상 세포주에서는 발현이 되지 않았으나 스트레스 유도제가 처리된 세포주에서는 ATFC의 전사체가 강하게 발현됨을 확인 할 수 있었다. 따라서 ATFC 유전자는 소포체 내의 정확하게 접혀지지 않았거나 조립되지 못한 단백질을 정확하게 접혀지게 하고 조립되게 하여 정상구조를 가지는 단백질로 재생하는 분자샤페론이나 folding enzyme의 전사를 촉진시키는 효모 Hac1p와 매우 유사한 기능을 수행할 것으로 추정할 수 있었다. 이상의 결과는, 효모를 제외한 모든 생물종에서 UPR pathway에 관련한 전사제어인자의 최초의 보고이다. 억제로 야기되는 것 같다.증진시키기 위해 행동변화단계에 따른 맞춤형 교육 프로그램을 개발하여 적용하였고, 그 결과, 행동변화단계별 교육 프로그램이 자궁경부암 조기검진의 수검 행동을 증진시키는데 효과적인 것으로 나타났다.lomus thermophilum, Thermotoga neapolitana 등에서 밝혀진 바와 같이 glutamic acid 부위가 xylanase의 활성부위라 여겨진다.倍), 수층(水層)이 약(約) 100배(100倍)로 나타나 홍삼(紅蔘)엑기스의 갈변색소형성(褐變色素形成)은 비효소적(非酵素的) 갈변반응(褐變反應)인 amino-carbonyl 반응(反應)이 주도적(主導的) 역할(役割)을 하고 있음을 알 수 있다. (6) 총당(總糖)과 갈변반응속도(褐變反應速度)는 유의성(有意性)이 있었으며 $100^{\circ}C$의 경우 20시간(20時間)에 가장 색도(色度)가 높아 갈변반응속도(褐變反應速度)가 0.2로 나타났다. the esophageal mucous cells pf Bryzoichthys lysimus contained small amount of neutral mucin, while on the other hand a feww mucous cells contained small amount of neutral mucin and minimal amount of sialomucin. But the esophageal mucous cells of Takifugu pardalis contained considerable amount of neutral mucin only.분해가 더욱 촉진되었으며, 30℃에서 교반 처리를 행한 경우가 10℃에서 교반 처리를 행한 경우 보다 지방분해가 더욱 촉진되었다. 산양유 원유는 30℃에서 교반 처리 시간이 연장되어도 지방분해는 뚜렷한 증가를 나타내지 않았다.와

• 생명과학회지
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• 제29권5호
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• pp.537-544
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• 2019

유전자 가위를 이용한 게놈편집 기술의 등장은 다양한 분야에서 분자육종에 대한 관심을 유발하였으며, 3세대 유전자가위 CRISPR 기술의 개발은 게놈편집을 통한 분자육종 시대를 가속화하고 있다. 본 연구에서는 최근 개발된 3세대 유전자 가위 CRISPR/Cas9을 이용하여 국내 보급품종인 백옥잠의 BmBLOS 유전자를 편집하여 돌연변이를 유도하고 유전형 및 표현형 검사를 통하여 유전자가위를 이용한 누에 분자육종가능성을 분석하고 이용기술을 확보하고자 하였다. 유전자 편집을 위하여 백옥잠의 BmBLOS 유전자의 염기서열을 구명하고, 이를 바탕으로 3종의 가이드 RNA를 합성하였다. 합성된 gRNA는 Cas9 단백질과 복합체를 형성시킨 후 BM-N 누에 세포주에 도입 후 T7 endonuclease I 분석을 통하여 편집효율이 가장 높은 B4N gRNA를 선발하였다. 누에 유전자를 편집하기 위하여 Cas9/B4N gRNA를 누에 초가 배아에 미세주사하고 사육하였다. 미세주사 후 부화율은 18% 가량으로 낮게 나타났으나 생존한 개체 중 돌연변이 발생율은 40% 이상으로 비교적 높게 나타났다. 또한 유전자 편집 G0 세대누에 중 70% 가량에서 표현형의 변화가 관찰되었고, 염기서열 분석결과 대부분의 개체에서 BmBLOS 유전자가 정상과 돌연변이가 같이 존재하는 이형접합자 형태로 나타났으며, 그 유전형 또한 모든 개체에서 다르게 나타났다. 이러한 결과에 비추어 볼 때 CRISPR/Cas9 시스템을 이용한 누에 분자육종의 가능성은 매우 높을 것으로 예상되나, 유전자 편집효율을 개선하고 동형접합자를 얻기 위한 교배 및 선발방법에 대한 지속적인 연구가 필요하다고 판단된다.

• Journal of Microbiology and Biotechnology
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• 제25권3호
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• pp.386-392
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• 2015

A novel recombinant bacmid, bEasyBm, that enables the easy and fast generation of pure recombinant baculovirus without any purification step was constructed. In bEasyBm, attR recombination sites were introduced to facilitate the generation of a recombinant viral genome by in vitro transposition. Moreover, the extracellular RNase gene from Bacillus amyloliquefaciens, barnase, was expressed under the control of the Cotesia plutellae bracovirus early promoter to negatively select against the nonrecombinant background. The bEasyBm bacmid could only replicate in host insect cells when the barnase gene was replaced with the gene of interest by in vitro transposition. When bEasyBm was transposed with pDualBac-EGFP, the resulting recombinant virus, EasyBm-EGFP, showed high levels of EGFP expression efficiency compared with that of non-purified recombinant virus BmGOZA-EGFP, which was constructed using the bBmGOZA system. In addition, nonrecombinant backgrounds were not detected in unpurified EasyBm-EGFP stocks. Based on these results, a high-throughput system for the generation of multiple recombinant viruses at a time was established.

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6351-6356
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• 2013

Background: Goshajinkigan (GJG) is used for the treatment of several neurological symptoms. We investigated the efficacy of GJG and mecobalamin (B12) against neurotoxicity associated with docetaxel (DOC) in breast cancer patients. Materials and Methods: Sixty breast cancer patients were treated with DOC. Thirty-three patients (GJG group) received oral administration of 7.5 g/day GJG and 27 patients (B12 group) received oral administration of 1500 ${\mu}g/day$ B12. Neuropathy was evaluated according to DEB-NTC (Neurotoxicity Criteria of Debiopharm), Common Terminology Criteria for Adverse Events (NCI-CTC) ver. 3.0, and a visual analogue scale (VAS). This study employed a randomized open design. Results: The incidence of neuropathy was 39.3% in the GJG group, and 88.9% in the B12 group (p<0.01). In the GJG group, grade 1 DEB-NTC was observed in 2 cases, grade 2 in 5 cases and grade 3 in 5 cases. Grade 1 NCI-CTC was observed in 7 cases, grade 2 in 6 cases, and VAS was $2.7{\pm}2.2$. In the B12 group, grades 1, 2 and 3 DEB-NTC were observed in one case, 12 cases and 12 cases, respectively; and grades 1, 2 and 3 NCI-CTC were observed in 11 cases, 12 cases and one case, and VAS was $4.9{\pm}2.4$. Conclusions: Concomitant administration of GJG is useful in preventing neuropathy in breast cancer patients treated with a DOC regimen.