• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.130-130
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• 2003

Gummy stem blight pathogen is very difficult not only to monitor the inoculum levels prior to host infection, and also it is destructive and hard to control in field condition. We have applied RAPD technique to elucidate the genetic diversity of the genomic DNA of Didymella bryoniae and also to generate specific diagnostic DNA probe useful for identification and detection. The 40 primers produced clear bands consistently from the genomic DNA of twenty isolates of Didymella bryoniae, and two hundred seventy-three amplified fragments were produced with 40 primers. The combined data from 273 bands was analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYS-PC (Version 1.80) to generate a dendrogram. At the distance level of 0.7, two major RAPD groups were differentiated among 20 strains. RAPD group (RG) I included 8 isolates from watermelon except one isolate from melon. RAPD group (RG) IV included 12 isolates from squash, cucumber, watermelon and melon.. In amplification experiment with SCAR specific primer RG1F-RG1R resulted in a single band of 650bp fragment only for 8 isolates out of 20 isolates that should be designated as RAPD Group 1. However, same set of experiment done with RGIIF-RGIIR did not result in any amplified product.. Our attempts to detect intraspecific diversity of ITS region of rDNA by amplifying ITS region and 17s rDNA region for 20 isolates and restriction digestion of amplified fragment with 12 enzymes did not reveal polymorphic band. In order to develop RAPD markers for RGIV specific primer, a candidate PCR fragment( ≒1.4kb) was purified and Southern hybridized to the amplified fragment RGIV isolates. This promising candidate probe recognized only RGIV isolates

• Fisheries and Aquatic Sciences
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• 제6권1호
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• pp.13-19
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• 2003

Genomic DNA from the muscle of marsh clam (Corbicula leana) from Gochang, Muan and a Chinese site was extracted to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction (RAPD- PCR). Out of 20 primers, seven primers produced amplified fragments which were consistently polymorphic. A total of 1,246 amplified products were produced of which 530 were polymorphic $(42.5\%)$. The number of polymorphic bands produced per primer varied from 40 to 122 with an average of 75.7 in marsh clam from Gochang. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic. Also, about $4.34\%$ of total polymorphic bands were specific to marsh clam from Gochang. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, which were polymorphic. This common bands in every individuals should be diagnostic of specific strains, species and/or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of 12 specific bands. The intra-population variation was revealed in the band patterns identified by this primer. The random primer OPB-12 (CCTTGACGCA) yielded the amplified fragments which were consistently polymorphic between the marsh clams from Gochang and from Muan. This primer produced a total of 77 polymorphic bands: 31 bands from Gochang, 14 from Muan and 32 from the Chinese populations. An average of polymorphic bands were 1.8 from Gochang and 2.5 from the Chinese populations. This value obtained from the Chinese population was higher than those from the two domestic populations. Generally, the RAPD polymorphism generated by these primers may be useful as a genetic marker for strain or population identification of marsh clam.

• Journal of Microbiology and Biotechnology
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• 제18권12호
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• pp.1938-1944
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• 2008

The procpb gene encoding human procarboxypeptidase B (proCPB, GeneBank access code AJ224866) was cloned and its Pichia expression plasmid, $pPIC9{\alpha}$/hproCPB (9.2 kb), was constructed, in which procpb was under the control of the AOXl promoter and connected to the downstream of the mating factor ${\alpha}$-1 ($MF{\alpha}1$) signal sequence. The plasmid was linearized by digestion with Sacl, and integrated into the genome of P. pastoris strain GS115. By culturing of Pichia transformant on methanol medium, the human proCPB was successfully expressed and secreted into the culture supernatant. Moreover, Western blot analysis of the extracellular proteins showed proCPB bands clearly at a molecular mass of 45 kDa, confirming the expression of proCPB with its right size. The CPB activity reached about 3.5 U/ml and 12.7 U/ml in the flask and fermentor batch cultures of Pichia transformant, respectively. No CPB enzyme activity was found in the intracellular fraction. When the fed-batch cultivation was performed with methanol and glycerol mixture as a feeding medium, the extracellular CPB activity was increased to 42.0 U/ml, which corresponds to a 3.3-fold higher level of CPB activity than that of batch culture. The $K_m$ and $k_{cat}$ values of recombinant human CPB enzyme for hippuryl-$_L$-Arg as a substrate were estimated to be 0.16 mM and $11.93\;sec^{-1}$, respectively.

• 한국미생물·생명공학회지
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• 제16권5호
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• pp.343-347
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• 1988

토양에서 분리한 알칼리 내성 Bacillus속의 chromosomal DNA로부터 promoter를 cloning하여 선별된 재조합 plasmid p-12내 의 promoter를 subcloning을 하였다. 그 결과 cloning된 promoter 내에는 서로 다른 두 가지의 promoter가 존재하는 것을 확인할 수 있었고 이로부터 각각의 promoter를 함유한 재조합 plasmid p-l2B1, p-l2B2를 제조하였다. 또한 CAT 비활성 측정에 의해 각 promoter의 활성을 비교해 본 결과 p-l2B1의 promoter는 p-l2B2의 promoter에 비해 상대적으로 높은 활성을 가지고 있었다. CAT 비활성을 생육시기에 따라 측정해 본 결과 p-l2B1과 p-l2B2는 대수증식기 이후 활성이 급증되었으며 배지 중 첨가된 1.0%의 glucose에 의해 활성이 억제되는 효과를 받았다.

• 한국미생물·생명공학회지
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• 제49권3호
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• pp.305-315
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• 2021

The cotton leafworm, Spodoptera littoralis, is a major pest in Egypt and many countries worldwide, and causes heavy economic losses. As a result, management measures to control the spread of the worm are required. S. littoralis nucleopolyhedrovirus (SpliNPV) is one of the most promising bioagents for the efficient control of insect pests. In this study, a chitinase gene (chitA) of a 1.8 kb DNA fragment was cloned and fully characterized from SpliNPV-EG1, an Egyptian isolate. A sequence of 601 amino acids was deduced when the gene was completely sequenced with a predicted molecular mass of 67 kDa for the preprotein. Transcriptional analyses using reverse transcription polymerase chain reaction (RT-PCR) revealed that chitA transcripts were detected first at 12 h post infection (hpi) and remained detectable until 168 hpi, suggesting their transcriptional regulation from a putative late promoter motif. In addition, quantitative analysis using quantitative RT-PCR showed a steady increase of 7.86-fold at 12 hpi in chitA transcription levels, which increased up to 71.4-fold at 120 hpi. An approximately 50 kDa protein fragment with chitinolytic activity was purified from ChitA-induced bacterial culture and detected by western blotting with an anti-recombinant SpliNPV chitinase antibody. Moreover, purification of the expressed ChitA recombinant protein showed in vitro growth inhibition of two different fungi species, Fusarium solani and F. oxysporum, confirming that the enzyme assembly and activity was correct. The results supported the potential role and application of the SpliNPV-ChitA protein as a synergistic agent in agricultural fungal and pest control programs.

• KSII Transactions on Internet and Information Systems (TIIS)
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• 제16권12호
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• pp.3904-3922
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• 2022

As a research hotspot, pedestrian detection has a wide range of applications in the field of computer vision in recent years. However, current pedestrian detection methods have problems such as insufficient detection accuracy and large models that are not suitable for large-scale deployment. In view of these problems mentioned above, a lightweight pedestrian detection and early warning method using a new model called you only look once (Yolov5) is proposed in this paper, which utilizing advantages of Yolov5s model to achieve accurate and fast pedestrian recognition. In addition, this paper also optimizes the loss function of the batch normalization (BN) layer. After sparsification, pruning and fine-tuning, got a lot of optimization, the size of the model on the edge of the computing power is lower equipment can be deployed. Finally, from the experimental data presented in this paper, under the training of the road pedestrian dataset that we collected and processed independently, the Yolov5s model has certain advantages in terms of precision and other indicators compared with traditional single shot multiBox detector (SSD) model and fast region-convolutional neural network (Fast R-CNN) model. After pruning and lightweight, the size of training model is greatly reduced without a significant reduction in accuracy, and the final precision reaches 87%, while the model size is reduced to 7,723 KB.

• 환경생물
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• 제17권4호
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• pp.439-448
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• 1999

1997년 11월부터 1998년 6월까지 총 4회에 걸쳐 군산인근 해역을 대상으로 Vibrio 속의 분포와 특성을 조사하였고, 분리동정된 Vibrio 속 3종(V. anguillarum, V. vulnificus, V, metschnikovii)을 대상으로 해수에서 수온 변화에 따른 생존을 관찰하였다. 조사 기간 중 총 해양 종속영양 세균의 분포는 평판도말법으로는 1.2$\pm$0.6$\times$$10^3$~2.0$\pm$1.5$\times$$10^4$CFU ml­$^1$을 나타냈으며, 형광현미경에 의한 직접측정법으로는 6.0$\pm$4.0$\times$$10^{5}$ ~1.9$\pm$1.5$\times$$10^{7}$ cells ml­$^1$의 범주에서 변화하여 측정방법에 따라 커다란 차이를 보였다. 해양 Vibrio의 분포는 1$\times$10~6$\pm$2.2$\times$$10^2$CFU ml­$^1$의 범주에서 변화하여 전반적으로 총 해양종속영양 세균의 수에 대하여 차지하는 비율은 0.l~6%였다. 최종 분리된 51균주를 Biolog Identification System$^{TM}$에 의해서 동정한 결과 V. mediterranei(11균주), V. anguillarum(13균주), V. netschnikovii(5균주), V. parahaemolyticus(5균주)등이 우점속으로 밝혀졌다. 분리 동정된 51균주들 간의 통계학적 유사도를 70%이상을 기준으로 grouping한 결과 26 group으로 나뉘어 본 조사해역에서 Vibrio 속의 다양성을 간접적으로 보여 주고 있다. 동정된 균주들에서 plasmid의 존재를 확인한 결과 65%에 해당하는 33균주가 plasmid를 갖고 있었으며 그 크기는 12kb이상으로 나타났다. 또한 분리된51균주들에 대하여 7종의 항생제 (gentamicin, ampicillin, chlorarnphenicol, streptomycin, kanamycin, tetracycline, carbenicillin)에 대한 내성을 측정한 결과 51균주들 중 96%가 한 종류 이상의 항세균제에 대하여 내성을 나타냈다. 분리 동정된 균주들 중 V. angulliarum, V. vulnificus, V. metschnikovii를 대상으로 여과된 해수에 접종시켜 생존율을 4, 15, $25^{\circ}C$에서 30일간 측정한 결과 15$^{\circ}C$에서 가장 높은 생존율을 보였다.

• 한국균학회지
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• 제41권4호
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• pp.268-273
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• 2013

흰가루병균(Podosphaera fusca)에 의한 오이 흰가루병의 생물적 방제를 위하여 검정한 길항세균 B. amyloliquefaciens M27 균주는 4.0% 흰가루병이 발생하였고 무처리는 80.5% 발생하였다. MH와 LB배지에서 배양한 B. amyloliquefaciens M27 균주의 배양여액은 TSB, NB와 KB배지에서 배양한 배양여액보다 오이 흰가루병 방제효과가 우수하였다. M27 균주를 LB배지에서 배양한 배양여액 2, 5, 10, 20, 50배와 100배 처리한 경우 오이 흰가루병이 0%, 0%, 0%, 1.3%, 3.1%, 5.0%와 33.3%를 나타낸 반면에 무처리는 60.0%를 나타내었다. 7월, 10월과 12월에 오이 흰가루병이 발생할 때 LB배지에서 배양한 M27균주의 배양여액를 10배로 처리한 결과 88.9~98.9% 방제효과를 나타났다. 이와 같은 결과에서 B. amyloliquefaciens M27 균주의 배양여액은 오이 흰가루병에 방제에 매우 효과적이었다.

• Asian-Australasian Journal of Animal Sciences
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• 제23권7호
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• pp.855-862
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• 2010

Among the known interferon-induced antiviral mechanisms, the Mx pathway is one of the most powerful pathways. The Mx protein has direct antiviral activity and inhibits a wide range of viruses by blocking an early stage of the viral replication cycle. Cloning, characterization, and expression of Mx in vivo and in vitro have been conducted. The chicken Mx gene spans 21 kb and is made up of 14 exons and 13 introns, of which the promoter region was analyzed. The real-time PCR results showed that Mx expression was increased in chicken embryo fibroblasts (CEF) after 12- and 24-h induction with polyI: C. Induction of Mx expression by poly I: C in vivo revealed tissue-specific patterns among the chicken tissues tested. A trace expression of Mx was detected in healthy chicken liver tissues from adult chickens without inducement; the expression levels in the liver, heart, and gizzard were higher than in the muscle and kidney. This is the first report to demonstrate the expression of a glutathione-S-transferase-tagged-Mx fusion protein of 75 KDa, as well as the biological activity tested by SDS-PAGE and western blotting.

• Journal of Microbiology and Biotechnology
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• 제12권3호
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• pp.389-397
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• 2002

Bacteriocin-producing lactic acid bacteria were isolated from kimchi. One isolate producing the most efficient bacteriocin was identified and named Lactococcus lactis B2, based on the biochemical properties and 16S rDNA sequences. The B2 bacteriocin inhibited many different Gram positive bacteria including Lactococcus, Lactobacillus, Leuconostoc, Enterococcus, Streptococcus, and Staphylococcus, but did not inhibit Gram-negative bacteria. The bacteriocin was maximally produced at temperatures between $25^{\circ}C\;and\;30^{\circ}C$ and at the initial pH of 7.0. Ninety $\%$ of the activity remained after 10 min of heat treatment at $121^{\circ}C,\;and\;100\%$, after 1 h exposure to organic solvents. The bacteriocin was purified from culture supernatant by ammonium sulfate precipitation, CM Sepharose column chromatography, ultrafiltration, and finally, by reverse-phase HPLC. A 1.58-kb fragment was amplified from B2 chromosome by using a primer set designed from the published nisA sequence. Sequencing result showed that the fragment contained the whole nisZ and 5' portion of nisB, whose gene product was involved in postmodification of nisin. The upstream sequence, however, was completely different from those of reported nisin genes.