• Journal of Plant Biology
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• 제39권1호
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• pp.31-40
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• 1996

Arabidopsis thaliana의 에틸렌 관련 돌연변이체인 에틸렌 과다 생성 돌연변이체(eto1-1)와 에틸렌에 대한 반응이 둔감한 돌연변이체(etr1-3과 ein2-1)의 생리·생화학적 특성을 분석하였다. 2∼3주된 Arabidopsis 식물로부터 얻은 성숙한 잎을 재료로 이용하였다. 에틸렌 생성량은 eto1-1은 야생형의 2배, etr1-3은 야생형의 4배, 그리고 ein2-1은 야생형의 4.5배 더 많았다. eto1-1에서의 ACC synthase와 ACC oxidase 활성은 야생형과 비슷하였으나 ACC 함량은 야생형보다 4.5배 더 많았다. ACC-N-malo-nyltransferase의 활성은 eto1-1이 야생형보다 3배 더 높았으며 SAM synthetase의 활성은 야생형보다 1.5배 더 높았다. 이들 결과로부터 eto1-1로의 변형은 SAM에서 ACC로 전환하는 과정 이전에 있음을 추측할 수 있다. etr1-3과 ein2-1에서 ACC synthase의 활성은 야생형보다 높았으나 ACC oxidase의 활성은 야생형보다 낮았다. 그러나 SAM synthetase 활성은 etr1-3에서는 야생형과 비슷하였고 ein2-1에서는 야생형보다 1.7배 높았다. 이것은 etr1-3과 ein2-1이 에틸렌에 대한 반응에 결함이 있기 때문에 그것으로 인하여 자가 조절이 되지 않았기 때문으로 추정된다. etr1-3의 ACC 함량은 야생형보다 2.3배 더 많았으며 ACC N-malonyltransferase의 활성은 야생형보다 3.9배 더 높았다. 그리고 ein2-1의 ACC 함량은 야생형보다 1.7배 더 많았으며 ACC N-malonyltransferase의 활성이 촉진된 것으로 추정된다. In vitro kinase assay를 한 결과 eto1-1과 ein2-1에서 36 kDa 단백질의 인산화를 관찰할 수 있었다.

• International Journal of Oral Biology
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• 제34권1호
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• pp.21-28
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• 2009

Mutations in DLX3 are associated with both autosomal dominant hypoplastic hypomaturation amelogenesis imperfecta (ADHHAI) and tricho-dento-osseous (TDO) syndrome. ADHHAI is caused by a c.561_562delCT (2bp-del DLX3) mutation whereas TDO syndrome is associated with a c.571_574delGGGG (4bp-del DLX3) mutation. However, although the causal relationships between DLX3 and an enamel phenotype have been established, the pathophysiological role of DLX3 mutations in enamel development has not yet been clarified. In our current study, we prepared expression vectors for wild type and deletion mutant DLX3 products (4bp-del DLX3, 2bp-del DLX3) and examined the effects of their overexpression on the expression of the enamel matrix proteins and proteases. Wild type DLX3 enhanced the expression of matrix metalloprotease 20 (MMP20) mRNA and protein in murine ameloblast-like cells. However, neither a 4bp-del nor 2bp-del DLX3 increased MMP20 expression. Wild type DLX3, but not the above DLX3 mutants, also increased the activity of reporters containing 1.5 kb or 0.5 kb of the MMP20 promoter. An examination of protein stability showed that the half-life of wild type DLX3 protein was less than 12 h whilst that of both deletion mutants was longer than 24 h. Endogenous Dlx3 was also found to be continuously expressed during ameloblast differentiation. Since inactivating mutations in the gene encoding MMP20 are associated with amelogenesis imperfecta, the inability of 4bp-del or 2bp-del DLX3 to induce MMP20 expression suggests a possible involvement of such mutations in the enamel phenotype associated with TDO syndrome or ADHHAI.

• Journal of Microbiology and Biotechnology
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• 제17권5호
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• pp.721-727
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• 2007

Stability-enhanced mutants, H44, 11-94, 5A2-84, and F8, of L-threonine aldolase(L-TA) from Streptomyces coelicolor A3(2)(SCO1085) were isolated by an error-prone PCR followed by a high-throughput screening. Each of these mutant, had a single amino acid substitution: H177Y in the H44 mutant, A169T in the 11-94 mutant, D104N in the 5A2-84 mutant and F18I in the F8 mutant. The residual L-TA activity of the wild-type L-TA after a heat treatment for 20 min at $60^{\circ}C$ was only 10.6%. However, those in the stability-enhanced mutants were 85.7% for the H44 mutant, 58.6% for the F8 mutant, 62.1% for the 5A2-84 mutant, and 67.6% for the 11-94 mutant. Although the half-life of the wild-type L-TA at $63^{\circ}C$ was 1.3 min, those of the mutant L-TAs were longer: 14.6 min for the H44 mutant, 3.7 min for the 11-94 mutant, 5.8 min for the 5A2-84 mutant, and 5.0 min for the F8 mutant. The specific activity did not change in most of the mutants, but it was decreased by 45% in the case of mutant F8. When the aldol condensation of glycine and 3,4-dihydroxybenzaldehyde was studied by using whole cells of Escherichia coli containing the wild-type L-TA gene, L-threo-3,4-dihydroxyphenylserine(L-threo-DOPS) was successfully synthesized with a yield of 2.0 mg/ml after 20 repeated batch reactions for 100 h. However, the L-threo-DOPS synthesizing activity of the enzyme decreased with increased cycles of the batch reactions. Compared with the wild-type L-TA, H44 L-TA kept its L-threo-DOPS synthesizing activity almost constant during the 20 repeated batch reactions for 100 h, yielding 4.0 mg/ml of L-threo-DOPS. This result showed that H44 L-TA is more effective than the wild-type L-TA for the mass production of L-threo-DOPS.

• BMB Reports
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• 제52권4호
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• pp.289-294
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• 2019

The present study evaluated the role of AHNAK in Bartonella henselae infection. Mice were intraperitoneally inoculated with $2{\times}10^8$ colony-forming units of B. henselae Houston-1 on day 0 and subsequently on day 10. Blood and tissue samples of the mice were collected 8 days after the final B. henselae injection. B. henselae infection in the liver of Ahnak-knockout and wild-type mice was confirmed by performing polymerase chain reaction, with Bartonella adhesion A as a marker. The proportion of B. henselae-infected cells increased in the liver of the Ahnak-knockout mice. Granulomatous lesions, inflammatory cytokine levels, and liver enzyme levels were also higher in the liver of the Ahnak-knockout mice than in the liver of the wild-type mice, indicating that Ahnak deletion accelerated B. henselae infection. The proportion of CD4+interferon-${\gamma}$ ($IFN-{\gamma}^+$) and $CD4^+$ interleukin $(IL)-4^+$ cells was significantly lower in the B. henselae-infected Ahnak-knockout mice than in the B. henselae-infected wild-type mice. In vitro stimulation with B. henselae significantly increased $IFN-{\gamma}$ and IL-4 secretion in the splenocytes obtained from the B. henselae-infected wild-type mice, but did not increase $IFN-{\gamma}$ and IL-4 secretion in the splenocytes obtained from the B. henselae-infected Ahnak-KO mice. In contrast, $IL-1{\alpha}$, $IL-1{\beta}$, IL-6, IL-10, RANTES, and tumor necrosis $factor-{\alpha}$ secretion was significantly elevated in the splenocytes obtained from both B. henselae-infected wild-type and Ahnak-knockout mice. These results indicate that Ahnak deletion promotes B. henselae infection. Impaired $IFN-{\gamma}$ and IL-4 secretion in the Ahnak-knockout mice suggests the impairment of Th1 and Th2 immunity in these mice.

• 생명과학회지
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• 제31권2호
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• pp.115-125
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• 2021

칼슘-의존성 단백질 카이네이즈(CDPK)는 식물의 Ca2+ 매개 신호 전달에 필수적인 역할을 한다. CDPK는 염분, 저온, 가뭄 등과 같은 비생물적 스트레스에 대한 식물의 반응을 조절하는 데 관여하는 것으로 알려져 있다. 벼의 CDPK는 31개의 유전자로 구성된 거대 다유전자군으로 되어 있지만, 단지 일부 벼의 CDPK 기능만이 확인되었다. 따라서, 벼에서 OsCPK11의 기능을 알아보기 위해, 이 연구는 염분, 저온 및 가뭄과 같은 비생물적 스트레스 조건에서 벼의 야생형과 ND0001 oscpk11 돌연변이체의 OsCPK11 발현 분석에 초점을 맞추었다. 염분, 저온, 가뭄 스트레스 처치를 위해 유식물을 각각 200 mM NaCl, 4℃, 20% PEG 6,000에 노출시켰다. 야생형과 ND0001 돌연변이체에서 OsCPK11의 발현을 확인하기 위해 RT-PCR과 quantitative real-time PCR을 수행하였다. RT-PCR 결과에 의하면, 야생형과 이형접합성 돌연변이체에서는 OsCPK11 전사체가 검출되었지만, 동형접합성 돌연변이체에서는 검출되지 않았다. Quantitative real-time PCR 결과에 의하면 야생형에서 염분, 저온, 가뭄 스트레스에 의해 OsCPK11의 상대적인 발현이 증가하였으며, 각각 24시간, 6시간, 24시간 후 최대 수준에 도달하였다. ND0001 동형접합성 돌연변이체의 OsCPK11의 상대적 발현은 야생형에 비해 현저히 감소하였다. 이러한 결과는 oscpk11 동형접합성 돌연변이체에서는 OsCPK11발현을 완전히 저해하며, OsCPK11유전자 발현 조절이 염분, 저온 및 가뭄 스트레스 신호 전달 과정에 관여할 수 있음을 의미한다.

• International Journal of Industrial Entomology and Biomaterials
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• 제33권2호
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• pp.113-120
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• 2016

Bombyx mandarina is widely accepted as ancestor of B. mori. Silkworms are served as well-characterized models for understanding the mechanism for the genetic regulation of development. In this study, we performed RNA-Seq analysis to examine tissue-expression of gloverin isoforms of the silk-gland, mid-gut, and fat body in B. mandarina. BLAST analysis revealed that four gloverin isoform gene sequences of B. mandarina were highly similar to B. mori. To identify the difference between two species, the expression profile of gloverin was measured by semi- RT-PCR analysis. The specific expression of gloverin isoform genes was observed mainly in the fat body from B. mori but not B. mandarina. However, all of tissues in the wild-type silkworm could induce the upregulation of compared with the B. mori. To validate the sudden increase in gloverin gene expression in the mid-gut tissue of B. mandarina, we were using qRT-PCR. Relative mRNA expression rate of gloverin at the wild-type silkworm was much higher than domestic silkworm. Comparative genomics between domesticated and wild silkworms showed different tissue-expression levels in some of immune related genes. These results are suggesting a trend toward decreasing immunity related genes expression during domestication. Further studies are needed to elucidate the silkworm domestication and an invaluable resource for wild silkworm genomics research.

• Parasites, Hosts and Diseases
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• 제51권4호
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• pp.441-448
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• 2013

This study describes the first record of Bourgelatia diducta (Nematoda: Chabertiidae) from wild boars in the Republic of Korea (=South Korea). Gastrointestinal tracts of 87 Korean wild boars (Sus scrofa coreanus) hunted in mountains in the south-western part of South Korea between 2009 and 2012 were examined for their visceral helminths. B. diducta, as identified by morphological characteristics of the head and tail, were recovered from the large intestine of 47 (54%) wild boars. The average length of adult female worms was $11.3{\pm}0.872$ mm and the thickest part of the body measured $0.54{\pm}0.04$ mm in maximum width, while those of males were $9.8{\pm}0.72$ and $0.45{\pm}0.03$ mm, respectively. The characteristic J-shaped type II ovejector was observed in females, and the type II dorsal ray with 2 rami on each side of the median fissure was uniquely seen in males. The buccal capsule was small, relatively thin-walled, cylindrical, very short, and ring-shaped. The externodorsal ray arose from a common stem with the dorsal ray. The cervical groove was absent. The anterior extremity was equipped with 20-22 external corona radiata, 4 cephalic papillae and 2 lateral amphids around the mouth. The eggs were $66.0{\times}38.9{\mu}m$ in average size. By the present study, B. diducta (Nematoda: Chabertiidae) is recorded for the first time in South Korea. Additionally, morphological characteristics and identification keys provided in the present study will be helpful in the faunistic or taxonomic studies for strongylid nematodes related.

• 미생물학회지
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• 제29권3호
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• pp.189-194
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• 1991

The degree of the genetic variations among Clostridium thermocellum ATCC 27405 and the wild type strains was investigated by the mehtod of GC ratio, DNA-DNA hybridization and RFLP (Restriction Fragment Length Polymorphism) patterns by ribosomal RNA and M13 probe. GC ratio and KNA homology values of th three isolates were approximately equal to those of ATCC type strain. The RFLP patterns by the rRNA and M13 probe showed some differences among C. thermocellum ATCC 27405, wild type strains and Clostridium thermohydrosulfuricum ATCC 33223, indicating that the two probes can be useful in subspecies- and apecies-identification.

• Journal of Pharmaceutical Investigation
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• 제39권5호
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• pp.345-351
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• 2009

Organic cation transporters (OCTs) are important for absorption, elimination of many endogenous small organic cations as well as a wide array of drugs and environmental toxins. This gene is located in a cluster on chromosome 6 and OCTs are in major organs such as intestine, liver, kidney, brain and placenta. Therefore, expression levels and function of OCTs directly affect plasma levels and intracellular concentrations of drugs and thereby determine therapeutic response. The aim of this study was to investigate the frequency of the SNPs on OCT1 (C181T and C1022T) and OCT2 (G808T) to analyze haplotype frequency in healthy Korean population. Human subjects have been genotyped for OCT1 (C181T for 195 subjects and C1022T for 825 subjects), using polymerase chain reaction-based diagnostic tests (RFLP). And for OCT2 (G808T), a total of 861 subjects have been genotyped, using pyrosequencing method. Haplotype was statistically inferred using an algorithm based on the expectation-maximization (EM). OCT1 C181T genotyping showed 100% homozygous wild-type (C/C). OCT1 C1022T genotyping showed wild-type (C/C), heterozygous (C/T) and homozygous mutant-type (T/T) and each accounted for 72.1, 24.5 and 3.4%, respectively. OCT2 G808T genotyping results also showed homozygous wild-type (G/G), heterozygous (G/T) and homozygous mutant-type (T/T) and each took 81.8, 17.9 and 0.3%, respectively. Based on these genotype data, haplotype analysis between OCT1 C181T and OCT1 C1022T has proceeded. The result has revealed that linkage disequilibrium between alleles is not obvious (P=0.0122).

• Journal of Forest and Environmental Science
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• 제30권1호
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• pp.161-166
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• 2014

To provide basic information for wildlife management, we investigated the distribution and ecology of wild mice in Mt. Keumwon recreational forests. The trapped wild mice were a total of 4 species such as Eothenomys regulus, Apodemus speciosus peninsulae, Apodemus agarius, and Crocidura suaveolens. A total of 79 individuals from four species were trapped. Number of Eothenomys regulus was the highest as 41 individuals followed by Apodemus speciosus peninsulae of 31 individuals, Apodemus agarius of 4 individuals, and Crocidura suaveolens of 4 individuals. Depending on forest type, 51 individuals were captured in broad-leaved forests, whereas 28 individuals were captured in coniferous forests. Average weight of the trapped Eothenomys regulus, Apodemus speciosus peninsulae, Apodemus agarius, and Crocidura suaveolens was 37.13 g, 36.29 g, 31.25 g, and 15 g, respectively. Among the trapped wild mice, vole which has the largest head and ears were Eothenomys regulus and vole which has the longest tail was Apodemus speciosus peninsulae and Apodemus agarius (p<0.5). This study might be valuable to control and predict some damages related to wild mice in Mt. Keumwon for protecting forest trees.