• Journal of Animal Science and Technology
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• 제49권4호
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• pp.437-442
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• 2007

본 연구는 축산과학원에서 보유중인 듀록 품종내에서 MC4R 유전자의 SNP 발현 특성과 육종가에 의한 선발 후 MC4R 유전자 빈도의 변화 및 경제형질에 대한 유전자형간 육종가의 차이를 구명하고자 수행하였다. 1999년부터 2005년까지 검정된 검정성적을 바탕으로 일당증체량, 등지방두께, 90kg 도달일령 및 사료요구율에 대하여 유전력과 유전상관 및 육종가를 추정하였으며, 2003년과 2004년에 출생한 660두에 대한 혈액을 채취하여 MC4R 유전자에 대한 유전자형 및 대립유전자 분석을 실시하였다. 또한 육종가에 의한 선발 후 유전자의 빈도변화를 세대별 그리고 선발군 및 도태군에 대하여 분석하였으며, 육종가의 분석결과를 이용하여 MC4R의 유전자형 효과를 보았다. 분석결과 육종가를 근거하여 선발한 MC4R 유전자형은 세대당 그리고 선발군과 도태군에서 차이를 보였으며, 각 형질별 육종가의 분산분석결과 사료요구율을 제외한 기타 경제형질에서 유의적으로 MC4R 유전자의 효과가 있는 것으로 나타났다. MC4R 유전자의 효과에 대한 보고는 상이한 부분도 있지만 자체 축군에 대한 다형성 분석 및 경제형질과의 연관성 분석에 의하여 축군 및 개량하고자 하는 경제형질에 따라 표지 유전인자로 활용하여 선발반응 및 정확도를 높일 수 있을 것으로 사료된다.

• Journal of Microbiology
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• 제44권5호
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• pp.530-536
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• 2006

Bacteriophage P4, a satellite phage of coliphage P2, is a very useful experimental tool for the study of viral capsid assembly and cos-cleavage. For an in vitro cos-cleavage reaction study of the P2-P4 system, new shortened and selectable markers containing P4 derivative plasm ids were designed as a substrate molecules. They were constructed by swapping the non-essential segment of P4 DNA for either the kanamycin resistance (kmr) gene or the ampicillin resistance (apr) gene. The size of the genomes of the resulting markers were 82% (P4 ash8 delRI:: kmr) and 79% (P4 ash8 delRI:: apr) of the wild type P4 genome. To determine the lower limit of genome size that could be packaged into the small P4-size bead, these shortened P4 plasmids were converted to phage particles with infection of the helper phage P2. The conversion of plasmid P4 derivatives to bacteriophage particles was verified by the heat stability test and the burst size determination experiment. CsCl buoyant equilibrium density gradient experiments confirmed not only the genome size of the viable phage form of shortened P4 derivatives, but also their packaging into the small P4-size head. P4 ash8 delRI:: apr turned out to be the smallest P4 genome that can be packaged into P4-sized head.

• Journal of Plant Biotechnology
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• 제7권4호
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• pp.267-272
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• 2005

Common bean (Phaseolus vulgaris L.) plants were regenerated via organogenesis from mature embryonic axes, cultured on MS medium supplemented with ildole-3-ecetic acid (IAA) and thidiazuron (TDZ) for one week in the dark. Embryonic axillary regions were excised, longitudinally cut to split the both sides, and cultured for two weeks on MS medium supplemented with IAA and TDZ. The combination 0.5 mg $l^{-1}$ TDZ/0.5 mg $l^{-1}$ IAA presented the higher efficiency in shoot regeneration and the combination 0.5 mg $l^{-1}$ TDZ/0.25 mg $l^{-1}$ IAA presented the higher efficiency in conversion of shoots to plants. Regenerating explants were transferred to MS medium containing 1 mg $l^{-1}$ BAP for shoot development. All elongated shoots were rooted in vitro, presented normal phenotype and produced viable seeds. Histological analysis confirmed the mode of regeneration as de novo shoot organogenesis.

• Journal of Microbiology and Biotechnology
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• 제25권5호
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• pp.696-703
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• 2015

A gamma-aminobutyric acid (GABA)-producing microorganism was isolated from jeot-gal (anchovy), a Korean fermented seafood. The isolate, A156, produced GABA profusely when incubated in MRS broth with monosodium glutamate (3% (w/v)) at 37℃ for 48 h. A156 was identified as Lactobacillus sakei by 16S rRNA gene sequencing. The GABA conversion yield was 86% as determined by GABase enzyme assay. The gadB gene encoding glutamate decarboxylase (GAD) was cloned by PCR. gadC encoding a glutamate/GABA antiporter was located immediately upstream of gadB. The operon structure of gadCB was confirmed by RT-PCR. gadB was overexpressed in Escherichia coli BL21(DE3) and recombinant GAD was purified. The purified GAD was 54.4 kDa in size by SDS-PAGE. Maximum GAD activity was observed at pH 5.0 and 55℃ and the activity was dependent on pyridoxal 5'-phosphate. The K_m and V_max of GAD were 0.045 mM and 0.011 mM/min, respectively, when glutamate was used as the substrate.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVII)
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• pp.243-246
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• 2005

A novel esterase showing high enantioselectivity to (S)-ketoprofen ethyl ester was selected from fosmid environmental DNA library which is provided by Microbial Genomic & Applications Center. As a result of Blast search, the gene wasn't registerated in Gene Bank yet. And as we know, conserved domain region of esterase , G-X-S-X-G, wasn't discovered.$^{4)}$ And it is similar to Beta-lactamase. The DNA sequence of cloned esterase include an open reading frame consisting of 1170 bp, designated as EST-Y29, encoding a protein of 389 amino acids with a molecular mass of about 42.8 kDa. And amino acid sequence analysis revealed only a few identity (28%) to tile known esterases/lipases in the databases containing the conserved sequence motifs of esterases/lipases. when being comparison to other esterase revealed , this enzyme seems to be classified as a new member of esterase family. EST-Y29 was functionally overexpressed in a soluble form in E. coli with maximum conversion yield of (S)-ketoprofen at $65^{\circ}C$. This study demonstrates that functional screening combined with the sequential uses of restriction enzymes to exclude already known enzymes is a useful approach for isolating novel enzyme from a metagenome.

• 한국양식학회지
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• 제16권1호
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• pp.51-58
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• 2003

The first Indian transgenic fish was generated in 1991 using borrowed constructs from foreign sources. To construct transformation vectors for the indigenous fishes, growth hormone genes of rohu (r-CH), Labeo rohita and catfish, Heteropneustes fossilis were isolated, cloned and sequenced; their fidelity was confirmed in prokaryotic and eukaryotic systems. A vector was constructed with grass carp b-actin promoter driving the expression of r-GH. Rohu eggs are large. fragile and swell 2~3 times. when fertilized. Hence they were amenable only for electroporated sperm-mediated gene transfer. Accordingly, the sperm electroporation technique was standardized to ensure 25% hatchling survival and 37% Presumptive transgenics without suffering any deformity. Southern analysis confirmed genomic integration in 15% of the tested individuals (Ti) belonging to family lines 2 and 3: another 25% of the Juveniles (Te) were also proved transgenic but with the transgene persisting extrachromosomally for longer than 1 to 2 years. perhaps due to the presence of replicon in the vector. Transgenics belonging to different family lines grew 6~8 times faster than the respective controls. Difference in growth trends of Ti and Te within a family line was not significant. In the Ti family 3 remarkable growth acceleration was sustained for a period longer than 36 weeks but in those of family 2, it gradually decreased. All transgenic fishes including the rohu converted the food at a significantly higher efficiency. Barring the transgenic mudloach, all the other transgenic fishes consumed food at significantly reduced rate.

• Journal of Microbiology and Biotechnology
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• 제21권3호
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• pp.277-283
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• 2011

Glyoxalase I catalyzes the conversion of methylglyoxal to S-D-lactoylglutathione in the presence of glutathione. The structural gene of glyoxalase I (GLO1) was cloned from an osmotolerant yeast, Candida magnoliae, which produces a functional sweetener, erythritol, from sucrose. DNA sequence analysis revealed that the uninterrupted open reading frame (ORF) of C. magnoliae GLO1 (CmGLO1) spans 945 bp, corresponding to 315 amino acid residues, and shares 45.2% amino acid sequence identity to Saccharomyces cerevisiae Glo1. The cloned ORF in a multicopy constitutive expression plasmid complemented the glo1 mutation of S. cerevisiae, confirming that it encodes Glo1 in C. magnoliae. The responses of CmGLO1 to environmental stresses were different from those of S. cerevisiae, which only responds to osmotic stress. An enzyme activity assay and reverse transcription polymerase chain reaction revealed that the expression of CmGLO1 is induced by stress inducers such as methylglyoxal, $H_2O_2$, KCl, and NaCl. The GenBank Accession No. for CmGLO1 is HM000001.

• Genomics & Informatics
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• 제2권2호
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• pp.86-91
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• 2004

Even though it represents $6-13\%$ of human genomic DNA, Alu sequences are rarely found in coding regions. When in exon region, over $80\%$ of them are found in 3' untranslated region (UTR). Pseudogenes are an important component of human genome. Their functions are not clearly known and the mechanism of how they are generated is still debatable. Both the Alu and Pseudogenes are important research problems in molecular biology. mRNA is thought to be a prime source of pseudogene and active research is going on its molecular mechanism. We report, for the first time, that mRNAs containing Alu repeats at 3' UTR has a significantly high correlation with processed pseudogenes, suggesting a possibility that Alu containing mRNAs have a high tendency to become processed pseudogenes. It is known that about $10\%$ of all human genes have been transposed. Transposed genes at 3' UTR without Alu repeat have about two processed pseudogenes per gene on average while we found with statistical significance that a transposed gene with Alu had over three processed Pseudogenes on average. Therefore, we propose Alu repeats as a new and important factor in the generation of pseudogenes.

• Journal of Microbiology and Biotechnology
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• 제29권9호
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• pp.1453-1459
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• 2019

Zeaxanthin is an important pigment in the photo-protection mechanism of microalgae. However, zeaxanthin epoxidase, an enzyme involved in the accumulation and conversion of zeaxanthin, has not been extensively studied in microalgae. In this work, we report the expression pattern of zeaxanthin epoxidase in Dunaliella tertiolecta (DtZEP) at different light and diverse salinity conditions. To confirm the responsiveness to light conditions, the ZEP expression pattern was investigated in photoperiodic (16 h of light and 8 h of dark) and continuous (24 h of light and 0 h of dark) light conditions. mRNA expression levels in photoperiodic conditions fluctuated along with the light/dark cycle, whereas those in continuous light remained unchanged. In varying salinity conditions, the highest mRNA and protein levels were detected in cells cultured in 1.5 M NaCl, and ZEP expression levels in cells shifted from 0.6 M NaCl to 1.5 M NaCl increased gradually. These results show that mRNA expression of DtZEP responds rapidly to the light/dark cycle or increased salinity, whereas changes in protein synthesis do not occur within a short period. Taken together, we show that DtZEP gene expression responds rapidly to light irradiation and hyperosmotic stress. In addition, ZEP expression patterns in light or salinity conditions are similar to those of higher plants, even though the habitat of D. tertiolecta is different.

• 한국미생물·생명공학회지
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• 제49권3호
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• pp.298-304
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• 2021

Photosynthetic conversion through cyanobacteria and microalgae is an increasingly serious concern in the global warming crisis. Many value-added substances are produced through strain improvement, and much research and development is being conducted to determine its potential as an actual industrial strain. Economic barriers throughout processing production can be overcome to produce value-added chemicals by microalgal strains. In this study, we engineered cyanobacteria strains for the photosynthetic production of squalene and confirmed the continuous cultivation of CO₂ and light conditions. The free-inducer system of gene expression was developed at the cyanobacterial strains. Then, the squalene production level and growth of the recombinant cyanobacteria were analyzed and discussed. For bio solar-cell factories, the ability to regulate genes based on the free-inducer gene expression system promotes metabolic engineering research and construction to produce value-added chemicals.