• Title/Summary/Keyword: host gene

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Platform of Hot Pepper Defense Genomics: Isolation of Pathogen Responsive Genes in Hot Pepper (Capsicum annuum L.) Non-Host Resistance Against Soybean Pustule Pathogen (Xanthomonas axonopodis pv. glycines)

  • Lee, Sang-Hyeob;Park, Do-Il
    • The Plant Pathology Journal
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    • v.20 no.1
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    • pp.46-51
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    • 2004
  • Host resistance is usually parasite-specific and is restricted to a particular pathogen races, and commonly is expressed against specific pathogen genotypes. In contrast, resistance shown by an entire plant species to a species of pathogen is known as non-host resistance. Therefore, non-host resistance is the more common and broad form of disease resistance exhibited by plants. As a first step to understand the mechanism of non-host plant defense, expressed sequence tags (EST) were generated from a hot pepper leaf cDNA library constructed from combined leaves collected at different time points after inoculation with non-host soybean pustule pathogen (Xanthomonas axonopodis pv. Glycines; Xag). To increase gene diversity, ESTs were also generated from cDNA libraries constructed from anthers and flower buds. Among a total of 10,061 ESTs, 8,525 were of sufficient quality to analyze further. Clustering analysis revealed that 55 % of all ESTs (4685) occurred only once. BLASTX analysis revealed that 74% of the ESTs had significant sequence similarity to known proteins present in the NCBI nr database. In addition, 1,265 ESTs were tentatively identified as being full-length cDNAs. Functional classification of the ESTs derived from pathogen-infected pepper leaves revealed that about 25% were disease- or defense-related genes. Furthermore, 323 (7%) ESTs were tentatively identified as being unique to hot pepper. This study represents the first analysis of sequence data from the hot pepper plant species. Although we focused on genes related to the plant defense response, our data will be useful for future comparative studies.

Molecular characterization and inhibition analysis of the acetylcholinesterase gene from the silkworm maggot, Exorista sorbillans

  • Lang, Guo-Jun;Zhang, Ming-Yan;Li, Bao-Ling;Yu, Lin-Lin;Lu, Xing-Meng;Zhang, Chuan-Xi
    • BMB Reports
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    • v.43 no.8
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    • pp.573-578
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    • 2010
  • Several organophosphorus (OP) insecticides can selectively kill the silkworm maggot, Exorista sorbillans (Es) (Diptera: Tachinidae), while not obviously affecting the host (Bombyx mori) larvae, but the mechanism is not yet clear. In this study, the cDNA encoding an acetylcholinesterase (AChE) from the field Es was isolated. One point mutation (Gly353Ala) was identified. The Es-353G AChE and Es-353A AChE were expressed in baculovirus-insect cell system, respectively. The inhibition results showed that for eserine and Chlorpyrifos, Es-353A AChE was significantly less sensitive than Es-353G AChE. Meanwhile, comparison of the I50 values of eserine, dichlorvos, Chlorpyrifos and omethoate of recombinant Es AChEs with its host (Bombyx mori) AChEs indicated that, both Es AChEs are more sensitive than B. mori AChEs. The results give an insight of the mechanism that some OP insecticides can selectively kills Es while without distinct effect on its host, B. mori.

Nigrospora Species Associated with Various Hosts from Shandong Peninsula, China

  • Hao, Yuanyuan;Aluthmuhandiram, Janith V.S.;Chethana, K.W. Thilini;Manawasinghe, Ishara S.;Li, Xinghong;Liu, Mei;Hyde, Kevin D.;Phillips, Alan J.L.;Zhang, Wei
    • Mycobiology
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    • v.48 no.3
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    • pp.169-183
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    • 2020
  • Nigrospora is a monophyletic genus belonging to Apiosporaceae. Species in this genus are phytopathogenic, endophytic, and saprobic on different hosts. In this study, leaf specimens with disease symptoms were collected from host plants from the Shandong Peninsula, China. The fungal taxa associated with these leaf spots were studied using morphology and phylogeny based on ITS, TEF1, and TUB2 gene regions. In this article, we report on the genus Nigrospora with N. gorlenkoana, N. oryzae, N. osmanthi, N. rubi, and N. sphaerica identified with 13 novel host associations including crops with economic importance such as bamboo and Chinese rose.

Bacillus thuringiensis 내에서 안정한 벡타를 이용한 cry1C 유전자의 발현

  • Choi, Soo-Keun;Oh, Keun-Hee;Kim, Jeong-Il;Park, Seung-Hwan
    • Microbiology and Biotechnology Letters
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    • v.25 no.6
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    • pp.566-570
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    • 1997
  • During sporulation, Bacillus thuringiensis strains produce crystals consist of toxin proteins highly specific against insect pests. Their host specificities are desirable from a standpoint of environmental safety, but also limit market potential. Thus, development of improved Bacillus thuringiensis strains having broad host spectrum will contribute to increase its use. For the construction of Bacillus thuringiensis strain having broad host spectrum, we cloned cry1C gene encoding a toxin protein highly toxic against Spodoptera exigua from a B. thuringiensis isolate and constructed two recombinant plasmids, pUBClC and plC60. The plasmid PUBC1C has a replication origin of the natural plasmid pBC16 from B. cereus which is closely related species to B. thuringiensis, and the pBC16 was known to be replicated by rolling-circle mechanism. The plasmid pIC60 has a replication origin of a resident 60 MDa plasmid from B. thuringiensis subsp. kurstaki HD263, and it is believed that the pIC60 is replicated in a theta mode. The two plasmids were introduced into B. thuringiensis subsp. kurstaki cryB strain, and the transformed strains produced well-shaped bipyramidal crystals. We confirmed the expression of the cry1C gene by SDS-PAGE, and Western blotting. By investigating the segregational stability, it was found that the plasmid pIC60 is more stable than the pUBC1C.

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Breeding of Aspergillus oryzae for the Alkaline Pretense Overproducing Strain. (재조합 Alkaline Protease를 대량 생산하는 Aspergillus oryzae 균주개발)

  • 이병로;유기원;최원균;최동성;임한진;성창근
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.450-455
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    • 1998
  • Aspergillus oryzae M-2-3 strain (argB$\^$-/) was transformed with pTAalp plasmid which was constructed for expression of the alkaline pretense gene, alpA, and 16 transformants were selected on arginine minus medium. When these transformants were tested for productivity of alkaline proteases using agar plate containing skim milk, the halo was observed around each colony of transformants, but not observed around the host strain in this condition. Southern analysis showed that the pTAalp plasmid having alpA gene was integrated into the chromosome of the host strain. The highest level of alkaline protease production was obtained in the culture filtrate of the transformant No. 14, which was estimated to 80-90% of total secreted proteins, and the enzyme activity was 64-450 times higher than those of host strain and industrial strain. Total nitrogen content and the digestion rate in soybean Koji extracts were also increased to 1.5 times in Aspergillus oryzae transformant No. 14.

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Molecular Cloning and Expression of Cellulase of Gene of Pseudomonas sp. in Escherichia coli (Pseudomonas sp.의 Cellulase 유전자의 대장균에의 클로닝 및 발현)

  • 정영철;김양우;노종수;성낙계;강신권
    • Microbiology and Biotechnology Letters
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    • v.18 no.6
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    • pp.633-639
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    • 1990
  • The genes for cellulases of Pseudomonas sp. LBC505 and CYC10, potent cellulase complex-producing strains, were cloned in Escherichia coli with pUC19. Recombinant plasmids pLCl and pLC2 were isolated from transformants producing cellulase by Congo red staining, and their genes cloned were 0.7 kb and 4.6 kb HindIII fragments, respectively. The inserts of pLCl and pLC2 were hybridized to chromosomal DNAs digested with HindIII from Pseudomona~ sp. LBC505 and CYC10, respectively. Immunodiffusion assays revealed that pLC1-and pLC2-encoded cellulase showed similarity with that of host strains. About 24% of cellulase activity was observed in the extracellular fraction of E. coli carrying pLC1, and its activity was higher about 1.4 times than that of LBC505. The enzymatic properties of pLC1 and pLC2 encoded cellulase were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases cloned were endocellulase.

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Hot Pepper Functional Genomics: Monitoring of Global Gene Expression Profiles During Non-Host Resistance Reactions in Hot Pepper Plant ( Capsicum annuum).

  • Lee, Sanghyeob;Chung, Eun-Joo;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.80.2-81
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    • 2003
  • Since hot peppers (Capsicum annuum L.) are getting reputation as an important source of vitamins, medicine and many other areas, consumption and cultivation is being increased in the world. In spite of this usefulness, so little attention has been given to the hot pepper plants. To date, less than 500 nucleotide sequences including redundancy has been identified in NCBI database. Therefore we started to EST sequencing project for initial characterization of the genome, because of the large genome size of hot pepper (2.7 3.3 ${\times}$ 109 bp), To date, a set of 10,000 non-redundant genes were identified by EST sequencing for microarray-based gene expression studies. At present, cDNA microarrays containing 4,685 unigene clones are used for hybridization labeled targets derived from pathogen infected and uninoculated leaf tissues. Monitoring of gene expression profiles of hot pepper interactions with soybean pustule pathogen (Xag;Xanthomonas axonopodis pv. glycine) will be presented.

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Stimulation of Actinorhodin Production by Streptomyces lividans with Chromosomally-Integrated Antibiotic Regulatory Gene, afsR2

  • Kim, Chang-Yeong;Park, Hyeon-Ju;Kim, Eung-Su
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.577-581
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    • 2003
  • Streptomyces lividans is one of the most commonly-used streptomyetes strain as a molecular cloning and expression host. Unlike its close relative S. coelicolor, however, S. lividans rarely produces secondary metabolite such as actinorhodin in a typical glucose-containing culture condition due to insufficient expression of some antibiotic regulatory genes including afsR2. Although multiple copies of afsR2 or a glycerol-specific culture condition stimulated actinorhodin production in S. lividans, both failed to stimulate actinorhodin production in S. lividans cultured in a typical glucose-containing medium. To generate a culture-condition-independent actinorhodin-overproducing S. lividans strain the afsR2 gene was integrated into the S. lividans TK21 chromosome via homologous recombination, followed by the genetic confirmation. This S. lividans strain produced a significant amount of actinorhodin in both glucose-containing liquid and plate cultures, with higher actinorhodin productivity compared to the S. lividans containing multiple copies of afsR2. These results suggest that a chromosomal integration of a single copy of an antibiotic regulatory gene is a promising method for the development of a stable antibiotic-overproducing streptomycetes strain.

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Research Trends in Bifidobacterium (비피더스 연구동향)

  • Park, Myeong-Soo;Ji, Geun-Eog
    • KSBB Journal
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    • v.25 no.4
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    • pp.319-329
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    • 2010
  • Bifidobacteria comprises up to 25% of the cultivable fecal bacteria in adults and 80% in infants. Many in vivo and clinical research results supporting its efficacy in the prevention and improvement of gastrointestinal health have been accumulated. As a consequence, expert committee WHO/FAO expert committee recommended Bifidobacterium as representative probiotics together with Lactobacillus acidophilus. In this review, research trends in bifidobacteria concerning the classification and identification of the genus Bifidobacterium, modulation of intestinal microflora, improvement of constipation, prevention of diarrhea, alleviation of atopy and allergy, barrier function through antimicrobial activity andimmune enhancement of the host will be introduced. Several gene expression systems based on bifidobacterial plasmids have been developed and successfully used to express several heterologous genes including anticancer proteins in Bifidogacterium. In animal test, bifidobacteria was proven to be a promising candidate for safe gene delivery system which can specifically colonize in the solid tumor.

The Bacteriophage λ DNA Replication Protein P Inhibits the oriC DNA- and ATP-binding Functions of the DNA Replication Initiator Protein DnaA of Escherichia coli

  • Datta, Indrani;Sau, Subrata;Sil, Alok Kumar;Mandal, Mitai C.
    • BMB Reports
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    • v.38 no.1
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    • pp.97-103
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    • 2005
  • Under the condition of expression of $\lambda$ P protein at lethal level, the oriC DNA-binding activity is significantly affected in wild-type E. coli but not in the rpl mutant. In purified system, the $\lambda$ P protein inhibits the binding of both oriC DNA and ATP to the wild-type DnaA protein but not to the rpl DnaA protein. We conclude that the $\lambda$ P protein inhibits the binding of oriC DNA and ATP to the wild-type DnaA protein, which causes the inhibition of host DNA synthesis initiation that ultimately leads to bacterial death. A possible beneficial effect of this interaction of $\lambda$ P protein with E. coli DNA initiator protein DnaA for phage DNA replication has been proposed.