• Journal of Microbiology and Biotechnology
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• 제25권11호
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• pp.1960-1965
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• 2015

Rabbit hemorrhagic disease virus (RHDV) is highly contagious and often causes fatal disease that affects both wild and domestic rabbits of the species Oryctolagus cuniculus. A highly pathogenic RHDV variant (RHDVa) has been circulation in the Korean rabbit population since 2007 and has a devastating effect on the rabbit industry in Korea. A highly pathogenic RHDVa was isolated from naturally infected rabbits, and the gene encoding the VP60 protein was cloned into a baculovirus transfer vector and expressed in insect cells. The hemagglutination titer of the Sf-9 cell lysate infected with recombinant VP60 baculovirus was 131,072 units/50 μl and of the supernatant 4,096 units/50 μl. Guinea pigs immunized twice intramuscularly with a trial inactivated RHDVa vaccine containing recombinant VP60 contained 2,152 hemagglutination inhibition (HI) geometric mean titers. The 8-week-old white rabbits inoculated with one vaccine dose were challenged with a lethal RHDVa 21 days later and showed 100% survival rates. The recombinant VP60 protein expressed in a baculovirus system induced high HI titers in guinea pigs and rendered complete protection, which led to the development of a novel inactivated RHDVa vaccine.

• BMB Reports
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• 제41권5호
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• pp.400-403
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• 2008

The insect baculovirus expression vector system (BEVS) is useful for producing biologically active recombinant proteins. However, the overexpressions of foreign proteins using this system often results in misfolded proteins and the formation of protein aggregates. To overcome this limitation, we developed a versatile baculovirus expression and secretion system using Bombyx mori protein disulfide isomerase (bPDI) as a fusion partner. bPDI gene fusion was found to improve the secretions and antibacterial activities of recombinant nuecin proteins. Thus, we conclude that bPDI gene fusion is a useful addition to BEVS for the large-scale production of bioactive recombinant proteins.

• The Plant Pathology Journal
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• 제29권3호
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• pp.330-337
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• 2013

Rice stripe virus (RSV), the type member of the genus Tenuivirus, transmits by the feeding behavior of small brown planthopper (SBPH), Laodelphax striatellus. To investigate the interactions between the virus and vector insect, total RNA was extracted from RSV-viruliferous SBPH (RVLS) and non-viruliferous SBPH (NVLS) adults to construct expressed sequence tag databases for comparative transcriptome analysis. Over 30 million bases were sequenced by 454 pyrosequencing to construct 1,538 and 953 of isotigs from the mRNA of RVLS and NVLS, respectively. The gene ontology (GO) analysis demonstrated that both libraries have similar GO structures, however, the gene expression pattern analysis revealed that 17.8% and 16.8% of isotigs were up- and down-regulated significantly in the RVLS, respectively. These RSV-dependently regulated genes possibly have important roles in the physiology of SBPH, transmission of RSV, and RSV and SBPH interaction.

• 농업과학연구
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• 제44권3호
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• pp.348-358
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• 2017

The green peach aphid, Myzus persicae, is one of the most serious insect pest and a vector for a multitude of viral diseases to many crops, vegetables, ornamentals, and fruit trees in the world. A large number of aphids can reduce plant vigor and cause defoliation. Many insecticides have been developed and applied to control the green peach aphid. However, this aphid has displayed a remarkable ability to establish resistance to almost every insecticide. We treated 5 different insecticides registered for M. persicae on pepper leaves and investigated the effects of the insecticides by measuring the time it took to achieve 90% control of the aphids. Acetamiprid worked faster than any other insecticides while cyantraniliprole showed the slowest insecticidal effect. Pymetrozine, pyrifluquinazon, and spirotetramet provided 90% control within similar time. Iwol population's control value was higher than any other populations 24 hours after treatment. When five different unregistered insecticides for M. persicae were treated on pepper leaves, no insecticidal effect was found for gamma-cyhalothrin and novaluron and spinosad showed an insecticidal effect of up to 70% in Iwol population only. Although chlorfenapyr and dinotefuran were not registered for M. persicae, their insecticidal effects were found to be 90% or higher.

• 농업과학연구
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• 제46권3호
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• pp.471-488
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• 2019

Random genes were screened in two transforming ways to investigate the new genes of a ladybug using the Harmonia axyridis cDNA library stock cell cloned in the LITMUS 28i vector in a previous study. Phenotypic variation was observed after injection of the synthesized double-stranded RNA through the in vitro transcription process. The cDNA library of H. axyridis was transformed into E. coli $DH5{\alpha}$ and 10B competent cells by heat shock. Analysis of the nucleotide sequences of the 42 clones with the insert DNAs revealed that 21 clones were homologous with the genes of insects, and only one clone had a gene from H. axyridis. Thirteen of the 21 insect genes were homologous with genes from coleopteran insects. Fourteen genes were selected, which were identified by the gene screening results, and were synthesized as double-stranded RNA through in vitro transcription. One microgram of the synthesized double-stranded RNA between segments T1 and T2 were injected using a syringe into each anesthetized fourth larvae which were under 2 days old. As a result, a phenotypic variation appeared in the larva injected with the two genes. While the eggs of H. axyridis injected with distilled water hatched out three days after oviposition, the eggs of H. axyridis injected with dsHma 06 did not hatch but become shrivel a week after oviposition. Most of the H. axyridis injected with dsHma 08 died and were unable to complete the pupation or eclosion during ecdysis.

• Safety and Health at Work
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• 제10권1호
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• pp.122-124
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• 2019

Simian malaria is a zoonotic disease caused by Plasmodium knowlesi infection. The common natural reservoir of the parasite is the macaque monkey and the vector is the Anopheles mosquito. Human cases of P. knowlesi infection has been reported in all South East Asian countries in the last decade, and it is currently the most common type of malaria seen in Malaysia and Brunei. Between 2007-2017, 73 cases of P. knowlesi infection were notified and confirmed to the Ministry of Health in Brunei. Of these, 15 cases (21%) were documented as work-related, and 28 other cases (38%) were classified as probably related to work (due to incomplete history). The occupations of those with probable and confirmed work related infections were border patrol officers, Armed Forces and security personnel, Department of Forestry officers, boatmen and researchers. The remaining cases classified as most likely not related to work were possibly acquired via peri-domestic transmission. The risk of this zoonotic infection extends to tourists and overseas visitors who have to travel to the jungle in the course of their work. It can be minimised with the recommended use of prophylaxis for those going on duty into the jungles, application of mosquito/insect repellants, and use of repellant impregnated uniforms and bed nets in jungle camp sites.

• 농약과학회지
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• 제13권4호
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• pp.309-314
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• 2009

2002년에서 2004년에 걸쳐 전국적으로 Rice stripe tenuivirus(RSV)에 의한 벼줄무늬잎마름병의 발생지역과 발생 포장율을 조사한 결과, RSV에 의한 발병은 점진적으로 감소하였다. RSV의 발생주율은 감수성 품종인 추청과 새추청, 일품에서 각각 45.8, 45.0 43.7%로 매우 높은 데 반해 저항성 품종인 화성에서는 4.4%로 낮았고, 좌지주율 역시 추청과 새추청, 일품 품종에서 각각 33.6, 33.2, 31.9%인데 비해 화성에서는 0.8%에 그쳤다. 두 가지 보독충 채집방법간의 효율성을 비교한 결과 기존의 포충망 채집에 비해 동력흡충기를 이용한 채집이 매우 효율적이었다. 2002년부터 2004년에 걸쳐 전국적으로 보독충 밀도를 조사한 결과, 연도별로 전국적인 평균 보독충율은 각각 3.6, 2.3, 1.3%로 시험기간에 걸쳐 보독충율은 전반적으로 감소하여 병 발생이 감소할 것으로 예측하였다. 이앙전 육묘상의 육묘상처리제 및 수면전개제 처리에 의한 애멸구 방제 효과를 조사한 결과, Imidacloprid 입제 등 4종 처리제의 약효가 인정되었다.

• 한국응용곤충학회지
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• 제54권2호
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• pp.91-97
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• 2015

담배가루이(Bemisia tabaci)는 외래해충으로 바이러스벡터로 작용하여, 토마토의 토마토황하잎말림병바이러스(TYLCV)를 비롯한 약 100여종의 바이러스를 매개하는 중요한 해충이다. 본 연구에서는 VIGS vector를 이용하여 담배가루이 방제를 위한 target 유전자들을 선발하기 위해 gateway system을 이용한 담배가루이 cDNA library 제작을 시도하였다. 첫 번째 방법으로 oligo d(T) primer를 사용하였을 때, 평균 약 1 kb의 insert와 $1.4{\times}10^4cfu$의 titer를 확인하였다. 그러나 insert size가 너무 커서 적절하지 않았다. 두 번째 방법으로 attB-N25 random primer를 이용하고, sonication을 6초 실시하여 다시 진행하였다. 그러나 확인되는 insert size는 다소 컸고, 몇몇은 insert가 너무 작아서 밴드가 확인 되지않았으며, $1.04{\times}10^54cfu$의 titer를 확인할 수 있었다. 세 번째 방법으로는 oligo d(T) primer를 이용하였고, sonication을 2초 실시하였다. 그 결과 300 bp~600 bp size의 insert가 확인되었으나, electro transformation을 사용한 첫번째, 두번째 방법에 비해 heat shock transformation을 사용하여 titer가 $5.2{\times}10^24cfu$로 매우 낮은 것을 확인 할 수 있었다. 결과적으로 cDNA library를 만들 때 먼저 random primer를 사용하여 First strand를 합성하여 poly A를 제거하고, 다음으로 sonication을 1초 실시하여 300~700 bp정도의 적절한 size의 insert를 생성하고, 마지막으로 electro-transformation을 실시하여 transformation 효율을 높인다면 VIGS vector에 적합한 cDNA library를 만들 수 있을 것으로 사료 된다.

• 한국산림과학회지
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• 제106권4호
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• pp.509-517
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• 2017

본 연구는 소나무재선충병의 확산 저지를 위해서 티아클로프리드 약제를 지상연막방제로 살포하였을 때 잣나무림 내 비표적 곤충 군집에 미치는 영향을 평가하기 위해 수행되었다. 또한 티아클로프리드의 살충 효과를 확인하기 위해서 표적 곤충인 북방수염하늘소와 대표적인 방화 곤충인 꿀벌에 대한 영향을 조사하였다. 2016년 강원대학교 학술림 내 잣나무림에서 처리구간(대조구와 약량별 처리구) 곤충류의 개체수, 종수, 종 구성의 유사도 그리고 곤충 2종의 치사율을 비교하였다. 각 조사구에는 곤충류 조사를 위해 6개의 함정트랩과 2개의 다중깔때기트랩을 설치하였고, 연무연막 살포 후 치사된 곤충류 조사를 위해 3장의 방수포를 배치하였다. 또한 북방수염하늘소와 꿀벌을 망실에 넣어 조사구 중심부에 위치한 잣나무의 7 m와 15 m 높이에 걸어 두었다. 조사 결과, 티아클로프리드를 1회 살포한 본 연구의 경우에는 곤충 군집이나 비표적 곤충인 꿀벌에 대한 영향은 미미한 것으로 확인되었다. 반면에 표적 곤충인 북방수염하늘소의 경우에는 티아클로프리드에 의한 치사 효과가 확인되었다. 망실 내 북방수염하늘소의 치사율은 망실의 높이와 공간적인 위치에 따라 달랐는데, 이는 티아클로프리드의 연무연막이 임내의 미기상 영향을 많이 받기 때문인 것으로 판단된다. 따라서 연무연막기를 이용하여 잣나무림 내 북방수염하늘소의 밀도와 확산을 억제하기 위해서는 연막기술을 개선함으로써 살충 효과에 대한 불확실성을 최소화시킬 필요가 있을 것이다.

• International Journal of Industrial Entomology and Biomaterials
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• 제18권1호
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• pp.18-27
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• 2009

B. thuringiensis strain LY-99 belonging to subsp. alesti (H3a3c), was isolated from Chinese tobacco warehouse and showed significantly high toxicity to Plutella xylostella. For the identification of the cry1-type genes from B. thuringiensis LY-99, an extended multiplex PCRrestriction fragment length polymorphism (PCRRFLP) method was established by using two pairs of universal primers based on the conserved regions of the cry1-type genes to amplify around 2.4 kb cry1-type gene fragments. Then the DNA fragment was cloned into pGEM-T Easy vector and digested with EcoRI and EcoRV enzymes. Through this method, a known cry1-type gene was successfully identified from the reference strain, B. thuringiensis subsp. alesti. In addition, the RFLP patterns revealed that B. thuringiensis LY-99 included a novel cry1A-type gene in addition to cry1Aa, cry1Ac, cry1Be and cry1Ea genes. The novel cry1A-type gene was designated cry1Ah2 (Genbank accession No DQ269474). An inverse PCR method was used to amplify the flank regions of cry1Ah2 gene. Finally, 3143 bp HindIII fragment from B. thuringiensis LY-99 plasmid DNA including 5' region and partial ORF was amplified, and sequence analysis revealed that cry1Ah2 gene from LY-99 showed 89.31% of maximum sequence similarity with cry1Ac1 crystal protein gene. In addition, the deduced amino acid sequence of Cry1Ah2 protein shared 87.80% of maximum identity with that of Cry1Ac2. This protein therefore belongs to a new class of B. thuringiensis crystal proteins.