Identification and Characterization of Baculoviruses isolated from Spodoptera litura

Yong Wang,Jae Young Choi,Jong Yul Roh,Qin Liu,Xueying Tao,Jong Bin Park,Soo Dong Woo,Byung Rae Jin,Yeon Ho Je 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.10

We isolated two baculoviruses, Spodoptera litura granulovirus (SlGV) and S. litura nucleopolyhedrovirus (SlNPV) in the dead larvae of S. litura. The granule of SlGV were ovoidal shape with an approximate measure of 240-340 nm×140-180 nm, and each granule contained one single rod-shape virion with a mean size of 180-200 nm×20-40 nm. Whereas, the polyhedra of SlNPV were irregular in shape with a approximate diameter of 1.0-1.5 ㎛, and numerous virions comprised of the multinucleocapsid were contained in each polyhedra. The major component of occlusion bodies produced by SlGV and SlNPV were about 29 and 30 kDa, respectively. When the phylogenic relationship between these viruses were analyzed using the nucleotide sequences of granulin gene from SlGV and polyhedrin gene from SlNPV, they were not closely related to each other. We also found that the two viruses showed similar insecticidal activity against 2nd instar larvae of Spodotera litura in terms of dose-response, but SlGV showed much longer LT50 than that of SlNPV. The two baculoviruses might be cooperatively be applied as biological control agent for the control of S. litura

Study on Interpretable Fuzzy Classification System Based on Neural Networks

Qin Yong,Xing Zong-yi,Jia Li-min,Wu Ying-ying 제어로봇시스템학회 2009 제어로봇시스템학회 국제학술대회 논문집 Vol.2009 No.8

This paper describes a comprehensive method to construct fuzzy classification system considering bothprecision and interpretability. Fuzzy classification system, initialized by modified Gath-Geva fuzzy clustering algorithm, is transformed into neural network. After training the neural network, fuzzy sets similarity measure is adopt to mergeredundant fuzzy sets to improve interpretability, and a constraint genetic algorithm is applied to improve precision. The simulation result on Iris data problem demonstrates the effectiveness of the proposed method

Molecular characterization of Bacillus thuringiensis serovar mogi (flagellar serotype 3a3b3d), a novel serogroup with mosquitocidal activity

Qin Liu,Jong Yul Roh,Yong Wang,Jae Young Choi,Xueying Tao,Jong Bin Park,Byung Rae Jin,Yeon Ho Je 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.10

The Bacillus thuringiensis strain K4 was isolated from fallen leaves, sampled in a forest region of the city of Mungyeong, Korea. The flagellated vegetative cells of B. thuringiensis strain K4 were agglutinated with the H3 reference antiserum and further, agglutinated with 3b and 3d monospecific antisera but non-reactive for 3c and 3e factor sera. These results create a new serogroup with flagellar antigenic structure of 3a3b3d, designated serovar mogi. The strain K4 showed high activity against dipteran larvae, Anopheles sinensis and Culex pipiens pallens while no lepidopteran toxicity. It produced a single ovoidal-shaped parasporal crystal whose SDS-PAGE protein profile consisted of several bands ranging from 75 to 30 kDa. Through the protein identification by nano-LC-ESI-IT MS analysis, the putative peptides of Cry19Ba, Cry40ORF2, Cry27Aa and Cry20Aa were detected. In contrast to the plasmid profile of B. thuringiensis H3 serotype strains, the strain K4 contained only a large plasmid (~100 kb) and we cloned partial cry27Aa, cry19Ba and cry40ORF2 genes from it by thermal asymmetric interlaced PCR. Sequencing analysis showed 87%, 88% and 88% homologous with known cry27Aa, cry19Ba and cry40ORF2 genes, respectively. The new type strain, B. thuringiensis subsp. mogi (H3a3b3d) will be a good resource for new mosquitocidal cry genes.

Effect of Structure Parameters and Wavelength on the Casein Concentration Detected by the Method of Scattered Transmitted Ratio

Yong Qin,Zhen Zhou,Lijie Wang 보안공학연구지원센터 2014 International Journal of Hybrid Information Techno Vol.7 No.3

Predicting connection integrity with consideration of thermo stress

Yong Qin Cheng,Shuo Qiong Liu,Feng Zhong Qi,Hui Ting Liu 대한기계학회 2022 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.36 No.8

In the conventional gas seal analysis of special threaded joints, the influence of temperature stress caused by the difference between formation temperature and fluid temperature in casing is often neglected. However, the influence mechanism of temperature stress is prerequisite information for accurately predicting the safe operating conditions that prevent a premium connection from being damaged. In this paper, we propose a model that includes these effects to predict the safe operating pressures and temperatures that will prevent seal failure. For the temperature stress, we propose a finite element model of thermal solid coupling. Subsequent stress changes are evaluated by a fully coupled model of thermal solid coupling to analyze the mechanical behavior of the premium connection. We predict the safe operating envelope (SOE) for premium connection failures caused by pressure and temperature perturbations after make-up. Our model predicts that radial pressure is a key factor for connection failure, especially for rapid temperature changes. If the formation temperature is high and the casing fluid temperature is low, the influence of temperature stress will increase under this working condition, resulting in an increase in the probability of failure of premium connections. Compared with conventional premium connection models, the model of thermal solid coupling predicts a smaller SOE when heating the internal casing fluid and a larger envelope when cooling the internal casing fluid. Finally, the heating rate was considered with respect to field applications. The heating rate was also considered, and slower heating/cooling rates can prevent damage to the seal integrity. Finally, the model was applied to explain several laboratory and field experiments and achieved good matches.

Characterization of a strain of Bacillus thuringiensis serovar aizawai which harbors a Rolling-Circle Replicating Plasmid, pBt1-3

Qin Liu,Jong Yul Roh,Yong Wang,Jae Young Choi,Xueying Tao,Jong Bin Park,Hee Jin Shim,Byung Rae Jin,Yeon Ho Je 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.05

Bacillus thuringiensis 1-3 (Bt 1-3) which was isolated from a Korean soil sample showed high insecticidal activity against Aedes aegypti as well as Plutella xylostella. The isolate was determined to belong to ssp. aizawai (H7) type by an H antiserum agglutination test and produced bipyramidal-shaped crystal proteins with a molecular weight of 130 kDa. PCR analysis with cry gene specific primers showed that Bt 1-3 contained cry1Aa, cry1Ab, cry1C, cry1D and cry2A gene, differing from spp. aizawai (reference strain) which contains cry1Aa, cry1Ab, cry1C and cry1D. We modified the plasmid capture system (PCS) to clone plasmid from Bt 1-3 through in vitro transposition. Fifty-three clones were acquired and their sizes were approximately 10 kb. Based on the sequence analysis, they were classified according to similarities with four known Bt plasmids, pGI3, pBMB175, pGI1 and pGI2, respectively. One of pGI3-like clones, named as pBt1-3, was fully sequenced and its 20 putative open reading frames (ORFs), Rep-protein, double-strand origin of replication (dso), single-strand origin of replication (sso), have been identified. The structure of pBt1-3 showed high similarity with pGI3 which is one of rolling-circle replication (RCR) group VI family.

Characterization of insecticidal Bacillus thuringiensis 1-3 strain and destruction of its sigG gene by homologous recombination

Qin Liu,Jong Yul Roh,Yong Wang,Hee Jin Shim,Jae Young Choi,Hong Guang Xu,Byung Rae Jin,Yeon Ho Je 한국응용곤충학회 2008 한국응용곤충학회 학술대회논문집 Vol.2008 No.05

A strain of Bacillus thuringiensis, named Bt 1-3, was isolated from Korean soil sample and it showed high insecticidal activity against Plutella xylostella. Bt 1-3 was deterimined to belong to ssp. aizawai (H7) by an H antiserum agglutination test and produced bipyramidal-shaped crystal proteins. PCR analysis with specific cry gene primers showed that Bt 1-3 contained cry1Aa, cry1Ab, cry1C, cry1D and cry2Ab genes. In addition, this isolate showed high uptake rate of foreign plasmid by electroporation. Based on these characteristics of Bt 1-3, we tried to construct a spore-free Bt 1-3 mutant by knock-out sigG gene, which is known as a key transcription factor during sporulation. First, we constructed a basal vector, named pDST, consisting of erythromycin resistant gene (EmR), partial polyhedrin gene and temperature sensitive origin of replication gene (Orits). Subsequently, according to the chromosomal DNA sequence of Bt subsp. konkukian 97-27, we amplifed upstream and downstream regions of Bt 1-3 sigG, and cloned into pDST (pDST-G). So far, several EmR colonies were obtained by electroporating into the wildtype Bt 1-3 and crossover by homologous recombination is going on.

A Novel Serogroup of Bacillus thuringiensis serovar mogi (flagellar serotype 3a3b3d) with Mosquitocidal Activity

Qin Liu,Yong Wang,Jae Young Choi,Xueying Tao,Jong Bin Park,Jae Su Kim,Yeon Ho Je 한국응용곤충학회 2011 한국응용곤충학회 학술대회논문집 Vol.2011 No.05

Bacillus thuringiensis (Bt) strain K4 was isolated from fallen leaves which had been collected at a forest stand in Mungyeong city, Republic of Korea. The flagellated vegetative cells of Bt K4 were agglutinated with the H3 reference antiserum among 55 reference H-antisera. In a further test to identify subfactors, 3b and 3d monospecific antisera were reactive to the cells, followed up with introducing a novel serogroup of 3a3b3d, designated as serovar mogi. The strain K4 had mosquitocidal activity against Dipteran larvae, Anopheles sinensis and Culex pipiens pallens, with no Lepidopteran toxicity observed. The SDS-PAGE profile of K4 crystal protein, ovoidal-shaped, included several bands ranging from 30-75 kDa. Four putative peptides, Cry19Ba, Cry40ORF2, Cry27Aa and Cry20Aa were detected from the bands by a nano-LC-ESI-IT MS analysis. Through a thermal asymmetric interlaced PCR, cry19Ba, cry40ORF2 and cry27Aa genes were partially cloned from K4 strain. Three cry genes were further found in the strain by a 454 pyrosequencing, ending up to showing 58%, 39% and 84% homology in amino acids with Cry56Aa, Cry8Ba and Cry39ORF2 toxins, respectively. This novel 3a3b3d type strain, B. thuringiensis subsp. mogi, can be used as a good resource for studying unknown mosquitocidal cry genes.

Cloning of B. thuringiensis Plasmids using a Modified Plasmid Capture System

Qin Liu,Jong Yul Roh,Yong Wang,Jae Young Choi,Hee Jin Shim,Hong Guang Xu,Xueying Tao,Byung Rae Jin,Yeon Ho Je 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.05

Bacillus thuringiensis 1-3 (Bt 1-3), belonging to subsp. aizawai (H7), showed different characteristics in plasmid profiles and had cry2A gene in addition to cry1Aa, cry1Ab, cry1C and cry1D. This strain exhibited dual insecticidal activity against Aedes aegypti as well as Plutella xylostella. Recently, we improved the donor-s of plasmid capture system (PCS) by inserting attB sites including lacZ between transposable elements (designated as pPCS-Troy), to construct E.coli-Bt shuttle vector. Through in vitro transposition with total plasmids DNA of Bt 1-3, 53 clones were acquired and their range of sizes were approximately 10 kb. Based on the sequence analysis, they were classified in 4 groups showing similarity with 4 known plasmids, pGI1, pGI2, pGI3 and pBMB175, respectively. One of pGI3-like clones was fully sequenced and its open reading frames were analyzed. As a donor for construction of shuttle vector, pDonr-attPEm vector harboring erythromycin resistant gene between attP sites was constructed. Through BP recombination with pPCS-Troy-cloned Bt plasmids and pDonr-attPEm, erythromycin resistant gene was transposed to Bt plasmids. This scheme proposes that in vitro transposition using pPCS-Troy and BP recombination using pDonr-attPEm can easily construct novel shuttle vectors with any Bt plasmids and this combined procedure can introduce foreign gengs into various circular DNA molecular.

Characterization of new B. thuringiensis plasmids and construction of E.coli-Bt shutter vectors using a modified plasmid capture system

Qin Liu,Jong Yul Roh,Yong Wang,Jae Young Choi,Xueying Tao,Hee Jin Shim,Byung Rae Jin,Yeon Ho Je 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.10

Bacillus thuringiensis 1-3 (Bt 1-3), belonging to subsp. aizawai (H7), showed different characteristics in plasmid profiles from type strain and had cry2A gene in addition to cry1Aa, cry1Ab, cry1C and cry1D. To clone its plasmids and construct E.coli-Bt shuttle vector, we constructed the plasmid capture system (PCS) by inserting attB sites including lacZ between transposable elements (designated as pPCS-Troy). Through in vitro transposition with total plasmids DNA of Bt 1-3, 53 clones were acquired and their sizes were approximately 10 kb. Based on the sequence analysis, they were classified in four groups showing similarities with four known Bt plasmids, pGI3, pBMB175, pGI1 and pGI2, respectively. One of pGI3-like clones, named as pBt1-3, was fully sequenced and its putative open reading frames (ORFs), Rep-protein, double-strand origin of replication (dso), single-strand origin of replication (sso), have been identified. The structure of pBt1-3 showed high similarity with pGI3 which is one of rolling-circle replication (RCR) group VI family. As a donor for construction of shuttle vector, pDonr-attPEm vector harboring erythromycin resistant gene between attP sites was constructed. Through BP recombination with pPCS-Troy-cloned Bt plasmids and pDonr-attPEm, erythromycin resistant gene was transposed to Bt plasmids. This scheme proposes that in vitro transposition using pPCS-Troy and BP recombination using pDonr-attPEm can easily clone Bt plasmids and construct novel shuttle vectors.