• 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.

• The Plant Pathology Journal
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• v.16 no.1
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• pp.29-35
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• 2000

The internal transcribed spacer regions (ITS I, 5.8S and ITS II) of the ribosomal DNAs were amplified from Korean isolates of Phytophthora spp. and sequenced to characterize them. Sequences from 33 isolates previously identified as P. boehmeriae, P. cactprum, P. cambivora, P. capsici, P. cinnamomi, P. erythroseptica, P. infestans, P. megasperma, P. melonis, P. nicotianae, P. palmivora and P. sojae were compared with published sequences, and a phylogenetic tree was produced. All isolates belonging to 10 species, P. cactorum, P. cambivora, P. capsici, P. cinnamomi P. citricola, P. infestans, P. nicotianae, P. palmivora and P. sojae were clearly clustered into published isolates of each species above 97% bootstrap value. Cucurbits isolates of Phytophthora previously identified as either P. melonis or P. drechsleri showed distinct evolutionary lineages from the P. megasperma was closely related to isolates of P. cryptogea-P. drechsleri showed distinct evolutionary lineages from the P. cryptogea-P. drechsleri complex group, indicating that P. melonis is a valid species. A Korean isolate of P. megasperma was closely related to isolates of P. erythroseptica showed distant genetic relationship with published isolates of P. erythroseptica (CBS 956.87). It is probable that the two Korean isolates could be genetically different from foreign isolates or misidentified. A grouping of species according to ITS sequence divergence matched, to some degree, the broad classification based on type of papilla. However, a separation of semi-papillate species and papillate species was not wvident in this study.

• The Plant Pathology Journal
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• v.21 no.1
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• pp.47-54
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• 2005

Identification of host genes involved in disease progresses and/or defense responses is one of the most critical steps leading to the elucidation of disease resistance mechanisms in plants. Soybean mosaic virus (SMV) is one of the most prevalent pathogen of soybean (Glycine max). Although the soybeans are placed one of many important crops, relatively little is known about defense mechanism. In order to obtain host genes involved in SMV disease progress and host defense especially for virus resistance, two different cloning strategies (DD RT-PCR and Subtractive hybridization) were employed to identify pathogenesis- and defenserelated genes (PRs and DRs) from susceptible (Geumjeong 1) and resistant (Geumjeong 2) cultivars against SMV strain G7H. Using these approaches, we obtained 570 genes that expressed differentially during SMV infection processes. Based upon sequence analyses, differentially expressed host genes were classified into five groups, i.e. metabolism, genetic information processing, environmental information processing, cellular processes and unclassified group. A total of 11 differentially expressed genes including protein kinase, transcription factor, other potential signaling components and resistant-like gene involved in host defense response were selected to further characterize and determine expression profiles of each selected gene. Functional characterization of these genes will likely facilitate the elucidation of defense signal transduction and biological function in SMV-infected soybean plants.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1539-1545
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• 2010

In beer, glutathione works as the main antioxidant compound, which also correlates with the stability of the beer flavor. In addition, high residual sugars in beer contribute to major nonvolatile components, which are reflected in a high caloric content. Therefore, in this study, the Saccharomyces cerevisiae GSH1 gene encoding glutamylcysteine synthetase and the Saccharomycopsis fibuligera ALP1 gene encoding ${\alpha}$-amylase were coexpressed in industrial brewing yeast strain Y31 targeting the ${\alpha}$-acetolactate synthase (AHAS) gene (ILV2) and alcohol dehydrogenase gene (ADH2), resulting in the new recombinant strain TY3. The glutathione content in the fermentation broth of TY3 increased to 43.83 mg/l as compared with 33.34 mg/l in the fermentation broth of Y31. The recombinant strain showed a high ${\alpha}$-amylase activity and utilized more than 46% of the starch as the sole carbon source after 5 days. European Brewery Convention tube fermentation tests comparing the fermentation broths of TY3 and Y31 showed that the flavor stability index for TY3 was 1.3-fold higher, whereas its residual sugar concentration was 76.8% lower. Owing to the interruption of the ILV2 gene and ADH2 gene, the contents of diacetyl and acetaldehyde as off-flavor compounds were reduced by 56.93% and 31.25%, respectively, when compared with the contents in the Y31 fermentation broth. In addition, since no drug-resistant genes were introduced to the new recombinant strain, it should be more suitable for use in the beer industry, owing to its better flavor stability and other beneficial characteristics.

• The Plant Pathology Journal
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• v.16 no.3
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• pp.142-146
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• 2000

Ordered differential display using RT-PCR (ODD-PCR) was conducted to have a profile of the differently expressed genes between a hypovirulent strain of Cryphonectria parasitica (UEP1) and its isogenic wild type strain (EP155/2). ODD-PCR has advantages of high sensitivity, reproducibility, proportional representation, and limited number of primer combinations comparing with other differential display methods. RNAs were prepared from 1 and 5 day liquid culture of both hypovirulent and wild type strains, and were further evaluated with the marker genes of C. parasitica such as cryparin and mating factor MF2-1, which were already proven to be specifically down-regulated by the presence of mycovirus CHV1-713. ODD-PCR was conducted using those RNAs and expressed genes were categorized to five groups according to their temporal and quantitative expression patterns. Those fives groups are CPC, CPE, CPL, CPD, and CPU which represent constitutively-expressed, early-expressed, late-expressed, down-regulated, and up-regulated, respectively. Ninety two primer combinations out of a total of 192 have been tested so far. Among the twenty to fifty distinct bands per each reaction, an average of four to ten genes was identified as viral-regulated fungal genes. Those viral-specifc genes were further analyzed by DNA sequencing followed by homology search. Characterization of 30 clones including all five groups were conducted as a preliminary data and more are under investigation.

• The Korean Journal of Mycology
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• v.25 no.3 s.82
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• pp.233-237
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• 1997

A PABA auxotroph of Pleurotus sajor-caju were transformed to prototrophy by using a plasmid containing pab 1 gene from Coprinus. The efficiencies of transformation of Pleurotus sajor-caju was five transformants per ${\mu}g$ of plasmid DNA. Southern blot analysis of DNA extracted from transformants demonstrated that plasmid DNA was integrated into the chromosomal DNA in multiple tandem copies. Progenies of heterokaryons between transformants of PABA and other auxotropic strains produced pab-progeny, which indicated that integration occurred at a site(s) other than the resident pab biosynthetic gene.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.425-429
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• 2009

Identification of virulent Bacillus cereus strains was examined by PCR using primers specific for the detection of plcR-papR, which encode regulatory proteins controlling the transcription of virulence factors in B. cereus. Total 96 strains of B. cereus that carried at least one of diarrheal toxin genes including hblACD, nheABC, and cytK showed all positive PCR products, while other 48 Bacillus strains that lacked the toxin genes were plcRpapR-negative. This PCR method targeting the plcR-papR genes appears to be simple and effective in identifying the enterotoxin genes-harboring B. cereus strains.

• Plant Breeding and Biotechnology
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• v.5 no.3
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• pp.237-242
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• 2017

AT-hook proteins of plant have shown to be involved in growth and development through the modification of chromatin architecture to co-regulate transcription of genes. Recently, many genes encoding AT-hook protein have been identified and their involvement in senescence delay is investigated. In this study, soybean transgenic plants overexpressing chromatin architecture-controlling ATPG7 gene was produced by Agrobacterium-mediated transformation and investigated for the positive effect on the important agronomic traits mainly focusing on yield-related components. A total of 27 transgenic soybean plants were produced from about 400 explants. $T_1$ seeds were harvested from all transgenic plants. In the analysis of genomic DNAs from soybean transformants, ATPG7 and Bar fragments were amplified as expected, 975 bp and 408 bp in size, respectively. And also exact gene expression was confirmed by reverse transcriptase-PCR (RT-PCR) from transgenic line #6, #7 and #8. In a field evaluation of yield components of ATPG7 transgenic plants ($T_3$), higher plant height, more of pod number and greater average total seed weight were observed with statistical significance. The results of this study indicate that the introduction of ATPG7 gene in soybean may have the positive effect on yield components.

• Genomics & Informatics
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• v.18 no.3
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• pp.27.1-27.12
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• 2020

The prevalence of early-onset type 2 diabetes (EOT2D) is increasing in Asian countries. Genome-wide association studies performed in European and various other populations have identified associations of numerous variants with type 2 diabetes in adults. However, the genetic component of EOT2D which is still unexplored could have similarities with late-onset type 2 diabetes. Here in the present study we aim to identify the association of variants with EOT2D in South Indian population. Twenty-five variants from 18 gene loci were genotyped in 1,188 EOT2D and 1,183 normal glucose tolerant subjects using the MassARRAY technology. We confirm the association of the HHEX variant rs1111875 with EOT2D in this South Indian population and also the association of CDKN2A/2B (rs7020996) and TCF7L2 (rs4506565) with EOT2D. Logistic regression analyses of the TCF7L2 variant rs4506565(A/T), showed that the heterozygous and homozygous carriers for allele 'T' have odds ratios of 1.47 (95% confidence interval [CI], 1.17 to 1.83; p = 0.001) and 1.65 (95% CI, 1.18 to 2.28; p = 0.006) respectively, relative to AA homozygote. For the HHEX variant rs1111875 (T/C), heterozygous and homozygous carriers for allele 'C' have odds ratios of 1.13 (95% CI, 0.91 to 1.42; p = 0.27) and 1.58 (95% CI, 1.17 to 2.12; p = 0.003) respectively, relative to the TT homozygote. For CDKN2A/2B variant rs7020996, the heterozygous and homozygous carriers of allele 'C' were protective with odds ratios of 0.65 (95% CI, 0.51 to 0.83; p = 0.0004) and 0.62 (95% CI, 0.27 to 1.39; p = 0.24) respectively, relative to TT homozygote. This is the first study to report on the association of HHEX variant rs1111875 with EOT2D in this population.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.11
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• pp.1519-1523
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• 2006

Through a screening and selection approach method, decamer random markers were used in a technique called random amplified polymorphic DNA (RAPD) assay with 252 genomic DNAs isolated from four major Haimen chicken populations: Rugao (62), Jiangchun (62), Wan-Nan (63) and Cshiqishi (65). A total of 3-score decamer random primers (S241-S260, S1081-S1100 and S1341-S1360) were employed in the preliminary RAPD-polymerase chain reaction (RAPD-PCR) assay with 50 random template DNA samples from all the populations. Four (6.67%) of the primers that produced obvious polymorphic patterns, interpretable and reproducible bands were selected and used with both the individual DNAs from each population and with pooled DNA samples of the four populations in subsequent analyses. The selected primers produced a total of 131 fragments with molecular size ranging from 835 to 4,972 base pairs (bp) when used with the individual DNAs; 105 (80.15%) of these fragments were polymorphic. With the pooled DNAs, 47 stable and characteristic bands with molecular size ranging from 840 to 4,983 bp, of which 23 (48.94%) polymorphic, were also generated. The band-sharing coefficient (BSC) calculated for the individuals in the population and among populations of bulked samples was between 0.8247 (Rugao) and 0.9500 (Cshiqishi); for pairwise populations, it was between 0.7273 (Rugao vs. Wan-Nan) and 0.9367 (Jiangchun vs. Cshiqishi) chicken populations. Using the BSC for individual and pairwise populations, the Nei's standard genetic distances between the chicken populations were determined and ranged from 0.0043 (Jiangchun vs. Cshiqishi) to 0.1375 (Rugao vs. Cshiqishi). The reconstructed dendrogram linked the Jiangchun and Cshiqishi chickens as closely related populations, followed by Wan-Nan, while the Rugao was the most genetically distant among the populations.