• Nutrition Research and Practice
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• 제16권1호
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• pp.14-32
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• 2022

BACKGROUND/OBJECTIVES: Peroxisome proliferator-activated receptor-gamma co-activator-1α (PGC-1α) has a central role in regulating muscle differentiation and mitochondrial metabolism. PGC-1α stimulates muscle growth and muscle fiber remodeling, concomitantly regulating lactate and lipid metabolism and promoting oxidative metabolism. Gynostemma pentaphyllum (Thumb.) has been widely employed as a traditional herbal medicine and possesses antioxidant, anti-obesity, anti-inflammatory, hypolipemic, hypoglycemic, and anticancer properties. We investigated whether G. pentaphyllum extract (GPE) and its active compound, gypenoside L (GL), affect muscle differentiation and mitochondrial metabolism via activation of the PGC-1α pathway in murine C2C12 myoblast cells. MATERIALS/METHODS: C2C12 cells were treated with GPE and GL, and quantitative reverse transcription polymerase chain reaction and western blot were used to analyze the mRNA and protein expression levels. Myh1 was determined using immunocytochemistry. Mitochondrial reactive oxygen species generation was measured using the 2'7'-dichlorofluorescein diacetate assay. RESULTS: GPE and GL promoted the differentiation of myoblasts into myotubes and elevated mRNA and protein expression levels of Myh1 (type IIx). GPE and GL also significantly increased the mRNA expression levels of the PGC-1α gene (Ppargc1a), lactate metabolism-regulatory genes (Esrra and Mct1), adipocyte-browning gene fibronectin type III domain-containing 5 gene (Fndc5), glycogen synthase gene (Gys), and lipid metabolism gene carnitine palmitoyltransferase 1b gene (Cpt1b). Moreover, GPE and GL induced the phosphorylation of AMP-activated protein kinase, p38, sirtuin1, and deacetylated PGC-1α. We also observed that treatment with GPE and GL significantly stimulated the expression of genes associated with the anti-oxidative stress response, such as Ucp2, Ucp3, Nrf2, and Sod2. CONCLUSIONS: The results indicated that GPE and GL enhance exercise performance by promoting myotube differentiation and mitochondrial metabolism through the upregulation of PGC-1α in C2C12 skeletal muscle.

• Parasites, Hosts and Diseases
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• 제57권2호
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• pp.161-166
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• 2019

This study was done to characterize distribution of Rickettsia spp. in ticks in the northwestern and southwestern provinces in the Republic of Korea. A total of 2,814 ticks were collected between May and September 2009. After pooling, 284 tick DNA samples were screened for a gene of Rickettsia-specific 17-kDa protein using nested PCR (nPCR), and produced 88 nPCR positive samples. Of these positives, 75% contained 190-kDa outer membrane protein gene (ompA), 50% 120-kDa outer membrane protein gene (ompB), and 64.7% gene D (sca4). The nPCR products of ompA, ompB, and sca4 genes revealed close relatedness to Rickettsia japonica, R. heilongjiangensis, and R. monacensis. Most Rickettsia species were detected in Haemaphysalis longicornis. This tick was found a dominant vector of rickettsiae in the study regions in the Republic of Korea.

• Genomics & Informatics
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• 제12권4호
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• pp.276-282
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• 2014

The disease tuberculosis, caused by Mycobacterium tuberculosis (MTB), remains a major cause of morbidity and mortality in developing countries. The evolution of drug-resistant tuberculosis causes a foremost threat to global health. Most drug-resistant MTB clinical strains are showing resistance to isoniazid and rifampicin (RIF), the frontline anti-tuberculosis drugs. Mutation in rpoB, the beta subunit of DNA-directed RNA polymerase of MTB, is reported to be a major cause of RIF resistance. Amongst mutations in the well-defined 81-base-pair central region of the rpoB gene, mutation at codon 450 (S450L) and 445 (H445Y) is mainly associated with RIF resistance. In this study, we modeled two resistant mutants of rpoB (S450L and H445Y) using Modeller9v10 and performed a docking analysis with RIF using AutoDock4.2 and compared the docking results of these mutants with the wild-type rpoB. The docking results revealed that RIF more effectively inhibited the wild-type rpoB with low binding energy than rpoB mutants. The rpoB mutants interacted with RIF with positive binding energy, revealing the incapableness of RIF inhibition and thus showing resistance. Subsequently, this was verified by molecular dynamics simulations. This in silico evidence may help us understand RIF resistance in rpoB mutant strains.

• 한국작물학회지
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• 제29권3호
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• pp.203-208
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• 1984

수도의 도열병 저항성 육종을 위하여 국제도열병 검정시험에서 여러 균계에 대해 고도의 안정된 저항성을 나타낸 Tadukan, Tetep 두 장간 저항성 품종을 6개 단간 리병성 계통들과 교배하여 도열병 저항성과 장간형질간의 연관관계를 분석하였는데 그 결과를 요약하면 다음과 같다. 1. Tadukan과 Tetep은 모든 단간계통들과의 조합에서 간장은 장간대 단간이 3 : 1로 분리되었으므로 이들은 각각 단순우성 장간 유전자를 지닌 것으로 나타났다. 2. Tadukan 품종은 T-2$^{+2}$ , C-8$^{+t}$ 균계에 대하여 도열병 저항성과 간장과는 연관이 없는 것으로 나타났다. 3. Tetep 품종도 역시 T-2$^{+1}$, C-8$^{+t}$ 균계에 대하여 도열병 저항성은 간장과는 연관이 없는 것으로 나타났다. 4. C-8$^{+t}$ 균계에 대한 Tadukan과 Tetep 품종의 저항성은 단순우성 유전자에 지배되고 있다. 5. T-2$^{+t}$ 균계에 대한 Tadukan과 Tetep 품종의 저항성은 각각 2개의 우성유전자에 의해 지배되고 이들은 서로 보족적인 관계에 있다.

• 한국미생물·생명공학회지
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• 제14권6호
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• pp.479-485
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• 1986

Bacillus amyloliquefaciens가 생성하는 액화형 $\alpha$-amylase의 구조유전자를 클로닝하기 위하여 이 균주 염색체의 2 - 5kb 획분을 분리하여 Mbo I으로 부분분해한 후 pUB110플라스미드의 BamHI 부위에 삽입하여 hybrid vector pSKS3 를 제작하였다. 이 재조합플라스미드를 세한수식결손 세포인 Bacillus subtilis ED 107에서 subcloning한 후 당화형 $\alpha$-amylase를 생성하는 Bacillus subtilis NA 64의 $\alpha$-amylase결실변이주 Bacills subtilis KM 107 주에서 형질 발현시켰다. 이때 형질전환빈도는 $10^{-5}$ - $10^{-7}$정도였으며 그중 약 50%가 $\alpha$-amylase 분비능이 있었으나 거의 대부분이 쉽게 실활되었다. 최종선별과정에서 $\alpha$-amylase를 가장 강하게 생성하는 형질전환주 Bacillus subtilis KM213으로부터 재 추출한 재조합플라스미드 pSKS3는 5.7kb 삽입된 단편이 BamH I/Mbo I 부위에 결합되어 있었다. pSKS3에 클로닝된 유전자에 의해 발현되는 $\alpha$-amylase 활성은 B. amyloliquefaciens 활성의 약 25%였으며, 이 pSKS3에 의해 발현되는 $\alpha$-amylase 단백은 B.amyloliquefaciens의 $\alpha$-amylase와 동일한 전기영동성을 나타내었음을 볼 때 pSKS3에 클로닝된 유전자는 B. amyloliquefaciens에서 유래함을 알 수 있었다.

• 한국작물학회지
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• 제44권4호
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• pp.345-349
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• 1999

This study was carried out to map Bphl and bph2 gene in Mudgo and Sangju13 (Oryza sativa L.) respectively conferring resistance to brown plan-thopper (BPH) and to establish the marker-assisted selection (MAS) system. Bulked seedling (grown for 20 days) test was conducted with the 73 F4 lines derived from a cross between Nagdongbyeo and Mudgo for Bphl and with 53 BC3F5 lines derived from the Milyang95/Sangju13 cross for bph2. Bph1 was mapped between RG413 and RG901 on chromo-some 12 at a distance of 7.5 cM from RG413 and 8.4 cM from RG90l. A recessive gene bph2 was located near RZ76 on chromosome 12 at a distance of 14.4 cM. Bphl and bph2 were linked to each other with a distance of about 30 cM. An RFLP marker, RG413 linked to Bphl, was converted to an STS marker to facilitate the marker-assisted selection. BPH resistant genotypes could be selected with 92% accuracy in a population derived from a line of NM47-B-B.

• 한국미생물·생명공학회지
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• 제49권1호
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• pp.75-87
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• 2021

Type I Interferons (IFNα) are known for their role as biological anticancer agents owing to their cell-apoptosis inducing properties. Development of an appropriate, cost-effective host expression system is crucial for meeting the increasing demand for proteins. Therefore, this study aims to develop codon-optimized IFNα-2b in L. lactis NZ3900. These cells express extracellular protein using the NICE system and Usp₄₅ signal peptide. To validate the mature form of the expressed protein, the recombinant IFNα-2b was screened in a human colorectal cancer cell line using the cytotoxicity assay. The IFNα-2b was successfully cloned into the pNZ8148 vector, thereby generating recombinant L. lactis pNZ8148-SP_Usp45-IFNα-2b. The computational analysis of codon-optimized IFNα-2b revealed no mutation and amino acid changes; additionally, the codon-optimized IFNα-2b showed 100% similarity with native human IFNα-2b, in the BLAST analysis. The partial size exclusion chromatography (SEC) of extracellular protein yielded a 19 kDa protein, which was further confirmed by its positive binding to anti-IFNα-2b in the western blot analysis. The crude protein and SEC-purified partial fraction showed IC₅₀ values of 33.22 ㎍/ml and 127.2 ㎍/ml, respectively, which indicated better activity than the metabolites of L. lactis NZ3900 (231.8 ㎍/ml). These values were also comparable with those of the regular anticancer drug tamoxifen (105.5 ㎍/ml). These results demonstrated L. lactis as a promising host system that functions by utilizing the pNZ8148 NICE system. Meanwhile, codon-optimized usage of the inserted gene increased the optimal protein expression levels, which could be beneficial for its large-scale production. Taken together, the recombinant L. lactis IFNα-2b is a potential alternative treatment for colorectal cancer. Furthermore, its activity was analyzed in the WiDr cell line, to assess its colorectal anticancer activities in vivo.

• 생명과학회지
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• 제20권7호
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• pp.1054-1065
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• 2010

생약복합물인 SH21B는 황금(Scutellaria baicalensis Georgi), 행인(Prunus armeniaca Maxim), 마황(Ephedra sinica Stapf), 석창포(Acorus gramineus Soland), 포황(Typha orientalis Presl), 원지(Polygala tenuifolia Willd), 하엽(Nelumbo nucifera Gaertner)의 혼합(비율 3:3:3:3:3:2:2)으로 이루어졌다. SH21B는 예로부터 한의학에서 비만의 치료에 사용되어 왔으나 자세한 분자적 메커니즘과 효능에 대한 연구는 이루어지지 않았다. 본 연구진은 선행연구를 통해 SH21B가 지방세포의 분화에서 adipogenesis (지방세포형성)와 관련된 유전자를 조절하여 중성지방의 축적을 억제함을 밝혔다. 본 연구에서는, microarray 기술을 이용하여 adipogenesis의 in vitro 모델인, 3T3-L1 세포에서 SH21B에 의한 지방세포형성 억제의 분자적 기작을 보다 상세하게 연구하고자 하였다. 전지방세포, 분화된 세포 그리고 SH21B에 의해 분화가 억제된 세포의 각각의 유전자 발현을 분석하기 위해 각 시료들에서 total RNA를 분리하여 cDNA를 합성한 후 microarray에 적용시켰다. 그 결과, 각각의 시료들의 비교에서 2배 이상의 유의한 발현 변화를 가지는 2,568개의 유전자를 확보하였다. 이 유전자들에 대해 Hierarchical clustering과 K-means clustering 분석을 진행하였고 서로 다른 양상을 가지는 9개의 군집(cluster)들을 분류하였다. 그 중, SH21B의 첨가에 의해 뚜렷하게 감소(cluster 4, cluster 6 및 cluster 9)하거나 반대로 뚜렷하게 증가(cluster 7와 cluster 8)하는 양상을 보이는 군집들을 따로 선별하여 그 군집들에 포함되어 있는 유전자들을 분석하였다. 선택 된 5개의 군집에는 지방세포형성과 세포증식에 관련된 유전자가 다수 포함되어 있었다. Cluster 4, cluster 6 그리고 cluster 9에는 peroxisome proliferator activated receptor gamma $\gamma$ ($PPAR{\gamma}$), CCAAT/enhancer binding protein $\alpha$ (C/$EBP{\alpha}$), sterol regulatory element binding transcription factor 1 (SREBF1), adiponectin (ADIPOQ), fatty acid synthase (FASN), lipoprotein lipase (LPL) 등의 지방세포형성 유도 및 관련 인자와 B-cell leukemia/lymphoma6 (BCL6), retinoblastoma 1 (RB1), cyclin-dependent kinase inhibitor 2C (CDKN2c), ras homolog gene family, member B (RHOB) 등의 많은 세포증식 억제 유전자가 포함되었다. 이와는 반대로, cluster 7과 cluster 8에는 $\beta$-catenin, cyclin D1 (CCND1), WNT1 inducible signaling pathway protein 2 (WISP2) 등과 같은 지방 세포형성 억제 조절자와 MARCKS-like1 (MARCKSL1), colony stimulating factor 1 (CSF1), discoidin domain receptor family, member 2 (DDR2), leukemia inhibitory factor receptor (LIFR) 등의 세포증식을 유도하는 조절자가 다수 포함되었다. 결론적으로, 이러한 결과들은 SH21B가 지방세포형성과 관련된 조절자 및 세포증식과 관련 된 조절자들의 유전자 발현을 조절하여 지방세포형성을 억제함을 제시한다.

• 약학회지
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• 제43권5호
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• pp.614-622
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• 1999

The standard operation procedure of mouse lymphoma L5178Y $tk^{+/-}-3.7.2C$ gene mutation assay (MOLY) has been regarded as a sensitive in vitro mammalian cell gene mutation assay that is capable of detecting clastogens as well as mutagens. Using MOLY, one of natural sweetner, stevioside (5mg/ml) and its aglycon, steviol ($340{\;}\mu\textrm{g}/ml$) were evaluated the mutagenicity. Stevioside and steviol did not induce mutagenicity in MOLY. On the other hand, stevioside (250mg/kg, B.W.) and steviol (200mg/kg, B.W.) were also evaluated their ability to induce micronuclei in regenerating hepatocytes and bone marrow cells of ddY mice. From these results, stevioside and steviol did not induce any mutagenic effect both MOLY and in vivo micronucleus test.

• Journal of Microbiology
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• 제42권2호
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• pp.111-116
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• 2004

It is known that Bacillus subtilis glutamyl-tRNA synthetase (GluRS) mischarges E. coli $tRNA_{1}$$^{Gln}$ with glutamate in vitro. It has also been established that the expression of B. subtilis GluRS in Escherichia coli results in the death of the host cell. To ascertain whether E. coli growth inhibition caused by B. subtilis GluRS synthesis is a consequence of Glu-$tRNA_{1}$$^{Gln}$ formation, we constructed an in vivo test system, in which B. subtilis GluRS gene expression is controlled by IPTG. Such a system permits the investigation of factors affecting E. coli growth. Expression of E. coli glutaminyl-tRNA synthetase (GlnRS) also amelio-rated growth inhibition, presumably by competitively preventing $tRNA_{1}$$^{Gln}$ misacylation. However, when amounts of up to 10 mM L-glutamine, the cognate amino acid for acylation of $tRNA_{1}$$^{Gln}$, were added to the growth medium, cell growth was unaffected. Overexpression of the B. subtilis gatCAB gene encoding Glu-$tRNA^{Gln}$ amidotransferase (Glu-AdT) rescued cells from toxic effects caused by the formation of the mis-charging GluRS. This result indicates that B. subtilis Glu-AdT recognizes the mischarged E. coli Glu-$tRNA_{1}$$^{Gln}$, and converts it to the cognate Gln-$tRNA_{1}$$^{Gln}$ species. B. subtilis GluRS-dependent Glu-$tRNA_{1}$$^{Gln}$ formation may cause growth inhibition in the transformed E. coli strain, possibly due to abnormal protein synthesis.