• Mycobiology
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• v.46 no.4
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• pp.429-439
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• 2018

To develop a convenient promoter analysis system for fungi, a null-pigment mutant (NPG) of Aspergillus nidulans was used with the 4'-phosphopantetheinyl transferase (PPTase) gene, npgA, which restores the normal pigmentation in A. nidulans, as a new reporter gene. The functional organization of serially deleted promoter regions of the A. nidulans trpC gene and the Cryphonectria parasitica crp gene in filamentous fungi was representatively investigated to establish a novel fungal promoter assay system that depends on color complementation of the NPG mutant with the PPTase npgA gene. Several promoter regions of the trpC and crp genes were fused to the npgA gene containing the 1,034-bp open reading frame and the 966-bp 3' downstream region from the TAA, and the constructed fusions were introduced into the NPG mutant in A. nidulans to evaluate color recovery due to the transcriptional activity of the sequence elements. Serial deletion of the trpC and crp promoter regions in this PPTase reporter assay system reaffirmed results in previous reports by using the fungal transformation step without a laborious verification process. This approach suggests a more rapid and convenient system than conventional analyses for fungal gene expression studies.

• Journal of Plant Biotechnology
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• v.41 no.4
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• pp.216-222
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• 2014

This study was carried out to develop an efficient transformation protocol via particle bombardment with PLBs (protocorm-like bodies) in Phalaenopsis. To achieve this aim, osmoticum treatment and an increasing shooting chances in particle bombardment process were applied for this study. In addition, pCAMBIA3301: ORE7 vector which contains a herbicide-resistance bar gene as a selectable marker and ORE7 gene as a gene of interests were employed. With regard to the increasing chances of shooting in particle bombardment, double shooting was the best results with 1.5 ~ 2.5 times higher than those of a single or triple shooting treatment in the productioon of PPT (D-L-phosphinothricin)-resistant PLBs. However, regeneration rate of shoots in double shooting was not high as a single shooting. Further, double shooting showed 35 ~ 40% higher than that of a single shooting in the frequency of browning. Regarding effects of different osmotic treatments, combination of 0.2 M sorbitol with 0.2 M mannitol showed the best results in transformation efficiency, regeneration of transformants and reduction of browning. Putative transgenic Phalaenopsis plants were analyzed by PCR analysis and confirmed the presence of bar and ORE 7 gene. Also, real-time PCR was conducted by using 21 transgenic plants and showed only 4 plants had one copy of transgene; whereas, the other 17 plants had more than 2 copies of transgene. Transgenic phalaenopsis plants produced in this study were transferred to pots and flowered normally without morphological variations in flower and leaf.

• Journal of Plant Biotechnology
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• v.33 no.1
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• pp.19-25
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• 2006

With the use of Agrobacterium and gene-gun, cold regulated gene (BN115) has been injected in Chrysanthemum leaf disc and transgenic plants have been produced successfully on the selection media containing phytohormone. To determine the presence of the transferred cold regulated gene (BN115) in the transgenic Chrysanthemum, PCR-amplification indicated the presence of that gene. Real-Time PCR for confirmation of the putative transgenic plants was established. The copy number of cold regulated gene (BN115) is extrapolated on the basis of a standard curve. Serial dilutions of known number of gene copies were in triplicates. In this diagram, PCR cycles are plotted against the fluorescence intensity. The cycle at which the fluorescence reaches a threshold cycle is inversely proportional to the starting amount of target DNA.

• 한국약용작물학회:학술대회논문집
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• 2007.05a
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• pp.253-254
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• 2007

• YAKHAK HOEJI
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• v.36 no.6
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• pp.582-590
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• 1992

S. pneumoniae recP mutant was compared with rec-8 mutant to identify whether they are the same gene or not by determining sensitivity to DNA damaging agents. recP and rec-8 mutant have almost same sensitivity to UV, ethylmethane sulfonate, and methylmethane sulfonate, suggesting that recP has the same function as the rec-8 gene in DNA repair.

• Journal of Korean Society of Industrial and Systems Engineering
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• v.19 no.38
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• pp.43-50
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• 1996

Evolution Programs, a form of Genetic Algorithms transformed from chromosome representation, are applied to the Timetabling of University which is one of the NP-hard problems. At the step of algorithms application, each class is established to be a specific category in feasible solution space. At. the same time, the exiting gene used in chromosome expression of Evolution Programs is modified to satisfy constraints effectively by transformation of gene which has multi-information. The new crossover method for fester operation in the Recombination attempted.. Roulette wheel selection and tournament selection are prepared.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2005.11a
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• pp.182-182
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• 2005

• Korean Journal of Microbiology
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• v.28 no.3
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• pp.181-187
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• 1990

STA gene coding glucoamylase was introduced into haploid Saccharomyces cerevisiae SHY3 and polyploid Saccharomyces cerevisiae 54. We constructed the recombinant plasmid by substituting the promoter region of alcohol dehydrogenase isoenzyme I gene for that of STA gene to increase the expression of STA gene and found that the activity of glucoamylase was increased in transformants. The plasmid stability was improved remarkably when we got the STA gene into the plasmid which had centromere. The activity of glucoamylase and transformation frequency of it, however, was decreased because of low copy number. Industrial polyploid strain was transformed with the recombinant plasmid having the $2\mu$ origin of replication and STA gene. It produced more alcohol than host when fermented in liquefied starch media. The industrial strain, however, was not transformed with the autonomously replicating plasmid containing centromere.

• Korean Journal of Environmental Biology
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• v.18 no.2
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• pp.205-213
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• 2000

Laboratory studies have revealed that naturally transformable bacteria develop competence under in situ conditions. Thus, the occurrence of competent bacteria in the environment can be considered as a certainty The persistence of free DNA in natural habitats is influenced by nucleolytic degradation and protection from degradation by adsorption to minerals. Although DNA seeded into natural environment was hydrolysed at substantial rates, but was still detectable at low levels after even several weeks. Compared to the number of laboratory based studies, only a few data have been published dealing with transformation of bacteria in the field. Recently, the potential transfer of recombinant DNA (rDNA) from deliberately or accidentally released bacteria to indigenous microbes has raised biosafety issues, since the persistence of rDNA becomes independent of the survival of its original host and leads to unpredictable, long-term ecological effects. The aim of the present review is to summarise recent literature on horizontal gene transfer (HGT) by transformation among bacteria in both soil and aquatic habitat and special emphasis is placed on recent reports which have addressed HGT among bacteria in the field. [Transformation, Horizontal gene transfer (HGT), recombinant DNA (rDNA), Genetically modified microorganisms (GMMs), Biosafety]

• Korean Journal of Microbiology
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• v.41 no.4
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• pp.239-245
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• 2005

Transformation-Associated Recombination (TAR) cloning technique allows selective isolation of chromosomal regions and genes from complex genomes. The procedure requires knowledge of relatively small genomic sequences that reside adjacent to the chromosomal region of interest. This technique involves homologous recombination during yeast spheroplast transformation between genomic DNA and a TAR vector that has 5' and 3' gene targeting sequences. In this study, we examined the effect of non-homologous spacing sequence in target hooks on homologous recombination using a plasmid model system. The efficiency of homologous recombination between the modified his3-TRP1-his3 fragments and HlS3 gene on plasmid were analyzed by the characterization of $Ura^+$ transformants. The numbers of $Ura^+$ transformant showed same level when seven different modified his3-TRP1-his3 fragments were used. But the percentage of positive recombinants. $Trp^+His^-$, dramatically decreased when used the modified his3-TRP1-his3 fragments contained incorrect spacing of nonhomologous region. As a result, we suggest that incorrect spacing inhibits the homologous recombination between target hook and substrate DNA. Therefore, we should consider the correct spacing in target hook when the target hook are used for cloning of orthologue gene.