• Title/Summary/Keyword: Gene structure

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Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionic Gonadotropin Receptor

  • Min, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.357-364
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$ -subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$-subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t631 or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632~653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agoinst-occupied receptors ~2- and ~17- fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Temporal variation in the community structure of green tide forming macroalgae(Chlorophyta; genus Ulva) on the coast of Jeju Island, Korea based on DNA barcoding (DNA 바코드를 이용한 제주도 연안 파래대발생(green tide)을 형성하는 갈파래(genus Ulva) 군집구조 및 주요 종 구성의 시간적 변이)

  • Hye Jin Park;Seo Yeon Byeon;Sang Rul Park;Hyuk Je Lee
    • Korean Journal of Environmental Biology
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    • v.40 no.4
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    • pp.464-476
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    • 2022
  • In recent years, macroalgal bloom occurs frequently in coastal oceans worldwide. It might be attributed to accelerating climate change. "Green tide" events caused by proliferation of green macroalgae (Ulva spp.) not only damage the local economy, but also harm coastal environments. These nuisance events have become common across several coastal regions of continents. In Korea, green tide incidences are readily seen throughout the year along the coastlines of Jeju Island, particularly the northeastern coast, since the 2000s. Ulva species are notorious to be difficult for morphology-based species identification due to their high degrees of phenotypic plasticity. In this study, to investigate temporal variation in Ulva community structure on Jeju Island between 2015 and 2020, chloroplast barcode tufA gene was sequenced and phylogenetically analyzed for 152 specimens from 24 sites. We found that Ulva ohnoi and Ulva pertusa known to be originated from subtropical regions were the most predominant all year round, suggesting that these two species contributed the most to local green tides in this region. While U. pertusa was relatively stable in frequency during 2015 to 2020, U. ohnoi increased 16% in frequency in 2020 (36.84%), which might be associated with rising sea surface temperature from which U. ohnoi could benefit. Two species (Ulva flexuosa, Ulva procera) of origins of Europe should be continuously monitored. The findings of this study provide valuable information and molecular genetic data of genus Ulva occurring in southern coasts of Korea, which will help mitigate negative influences of green tide events on Korea coast.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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Genetic Relationship between Populations and Analysis of Genetic Structure in Hanwoo Proven and Regional Area Populations (한우 종모우와 지역별 한우 집단의 유연관계와 유전적 구조 분석)

  • Oh, Jae-Don;Jeon, Gwang-Joo;Lee, Hak-Kyo;Cho, Byung-Wook;Lee, Mi-Rang;Kon, Hong-Sik
    • Journal of Life Science
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    • v.18 no.10
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    • pp.1442-1446
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    • 2008
  • Seven populations of 586 Hanwoo have been characterized by using 10 microsatellite DNA markers. Size of microsatellite markers decided using GeneMapper Software (v.4.0) after analyze in kinds of ABI machine of name of 3130. Frequencies of microsatellites markers were used to estimate heterozygosities and genetic distances. Genetic distancesbetween populations were obtained using Ne's DA distance method. Expected heterozygosity between each population was estimated very analogously. Genetic distances (0.0413) between Kangwan (KW) and Gyonggi (GG), Jeonpuk (JP) were nearest than distances between other populations by 0.021. Genetic distances between Gyonggi (GG) and Kyongpuk (KP) showed far distance than other populations by 0.032. In the UPGMA tree that is made based on DA distance matrix. Each individuals were not ramified to different group and were spread evenly in phylogenetic dendrogram about all Hanwoo of each regional area populations. But Hanwoo proven population was ramified to different group.

Genetic Relationship between Populations and Analysis of Genetic Structure in the Korean Native Chicken and the Endemic Chicken Breeds (한국재래닭 및 토착화 품종간의 유연 관계 및 유전 특성 분석)

  • Oh, J.D.;Kang, B.S.;Kim, H.K.;Park, M.N.;Chae, E.J.;Seo, O.S.;Lee, H.K.;Jeon, G.J.;Kong, H.S.
    • Korean Journal of Poultry Science
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    • v.35 no.4
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    • pp.361-366
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    • 2009
  • The purpose of this study was to assess the genetic variation and establish the relationship amongst breeds and strains using 7 chicken specific microsatellite markers. A total of 317 DNA samples from four Korean native chicken (KNC) strains (KR: Korean Native Red chicken strain, KY: Korean Native Yellow chicken strain, KL: Korean Native Black chicken strain, KO: Ogol chicken strain) and three introduced endemic chicken breeds (LE: Leghorn chicken breed, RI: Rhode Island Red chicken breed, CO: Cornish chicken breed). The size of microsatellite markers was decided using GeneMapper Software (v.4.0) after being analyzed using an ABI 3130 Genetic Analyzer. Frequencies of microsatellites markers were used to estimate heterozygosities and genetic distances. The lowest distance (0.074) was observed between the KY and KL breeds and the highest distance (0.779) between the KL and LE breeds. The KNC strains (KR, KY, KL) have comparatively near genetic distance each other. On the other side, each individual was not ramified to different groups and were spread evenly in phylogenetic dendrogram about all the KNC of each strain populations. But the endemic breed populations (LE, RI, CO) were ramified to different groups. The microsatellite polymorphism data were shown to be useful for assessing the genetic relationship between Korean native strains and other foreign breeds.

Current status of Brassica A genome analysis (Brassica A genome의 최근 연구 동향)

  • Choi, Su-Ryun;Kwon, Soo-Jin
    • Journal of Plant Biotechnology
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    • v.39 no.1
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    • pp.33-48
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    • 2012
  • As a scientific curiosity to understand the structure and the function of crops and experimental efforts to apply it to plant breeding, genetic maps have been constructed in various crops. Especially, in the case of Brassica crop, genetic mapping has been accelerated since genetic information of model plant $Arabidopsis$ was available. As a result, the whole $B.$ $rapa$ genome (A genome) sequencing has recently been done. The genome sequences offer opportunities to develop molecular markers for genetic analysis in $Brassica$ crops. RFLP markers are widely used as the basis for genetic map construction, but detection system is inefficiency. The technical efficiency and analysis speed of the PCR-based markers become more preferable for many form of $Brassica$ genome study. The massive sequence informative markers such as SSR, SNP and InDels are also available to increase the density of markers for high-resolution genetic analysis. The high density maps are invaluable resources for QTLs analysis, marker assisted selection (MAS), map-based cloning and comparative analysis within $Brassica$ as well as related crop species. Additionally, the advents of new technology, next-generation technique, have served as a momentum for molecular breeding. Here we summarize genetic and genomic resources and suggest their applications for the molecular breeding in $Brassica$ crop.

In vivo Antifungal Activity of Pyrrolnitrin Isolated from Burkholderia capacia EB215 with Antagonistic Activity Towards Colletotrichum Species (탄저병균에 대하여 길항작용을 보이는 Burkholderia cepacia EB215로부터 분리한 Pyrrolnitrin의 항균활성)

  • Park, Ji-Hyun;Choi, Gyung-Ja;Lee, Seon-Woo;Jang, Kyoung-Soo;Choi, Yong-Ho;Chung, Young-Ryun;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Mycology
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    • v.32 no.1
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    • pp.31-38
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    • 2004
  • An endophytic bacterial strain EB215 that was isolated from cucumber (Cucumis sativus) roots displayed a potent in vivo antifungal activity against Colletotrichum species. The strain was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, and 16S rDNA gene sequence. Optimal medium and incubation period for the production of antifungal substances by B. cepacia EB215 were nutrient broth (NB) and 3 days, respectively. An antifungal substance was isolated from the NB cultures of B. cepacia EB215 strain by centrifugation, n-hexane partitioning, silica gel column chromatography, preparative TLC, and in vitro bioassay. Its chemical structure was determined to be pyrrolnitrin by mass and NMR spectral analyses. Pyrrolnitrin showed potent disease control efficacy of more than 90% against pepper anthracnose (Colletotrichum coccodes), cucumber anthracnose (Colletotrichum orbiculare), rice blast (Magnaporthe grisea) and rice sheath blight (Corticium sasaki) even at a low concentration of $11.1\;{\mu}g/ml$. In addition, it effectively controlled the development of tomato gray mold (Botrytis cinerea) and wheat leaf rust (Puccinia recondita) at concentrations over $33.3\;{\mu}g/ml$. However, it had no antifungal activity against Phytophthora infestans on tomato plants. Further studies on the development of microbial fungicide using B. cepacia EB215 are in progress.

Analysis of Global Gene Expression Profile of Human Adipose Tissue Derived Mesenchymal Stem Cell Cultured with Cancer Cells (암세포주와 공동 배양된 인간 지방 조직 유래 중간엽 줄기 세포의 유전자 발현 분석)

  • Kim, Jong-Myung;Yu, Ji-Min;Bae, Yong-Chan;Jung, Jin-Sup
    • Journal of Life Science
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    • v.21 no.5
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    • pp.631-646
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    • 2011
  • Mesenchymal stem cells (MSC) are multipotent and can be isolated from diverse human tissues including bone marrow, fat, placenta, dental pulp, synovium, tonsil, and the thymus. They function as regulators of tissue homeostasis. Because of their various advantages such as plasticity, easy isolation and manipulation, chemotaxis to cancer, and immune regulatory function, MSCs have been considered to be a potent cell source for regenerative medicine, cancer treatment and other cell based therapy such as GVHD. However, relating to its supportive feature for surrounding cell and tissue, it has been frequently reported that MSCs accelerate tumor growth by modulating cancer microenvironment through promoting angiogenesis, secreting growth factors, and suppressing anti-tumorigenic immune reaction. Thus, clinical application of MSCs has been limited. To understand the underlying mechanism which modulates MSCs to function as tumor supportive cells, we co-cultured human adipose tissue derived mesenchymal stem cells (ASC) with cancer cell lines H460 and U87MG. Then, expression data of ASCs co-cultured with cancer cells and cultured alone were obtained via microarray. Comparative expression analysis was carried out using DAVID (Database for Annotation, Visualization and Integrated Discovery) and PANTHER (Protein ANalysis THrough Evolutionary Relationships) in divers aspects including biological process, molecular function, cellular component, protein class, disease, tissue expression, and signal pathway. We found that cancer cells alter the expression profile of MSCs to cancer associated fibroblast like cells by modulating its energy metabolism, stemness, cell structure components, and paracrine effect in a variety of levels. These findings will improve the clinical efficacy and safety of MSCs based cell therapy.

Ultrastructural analysis and quantification of autophagic vacuoles in wild-type and atg5 knockout mouse embryonic fibroblast cells (정상 및 atg5 유전자 제거 섬유아세포에서 자가포식체의 미세구조 및 이들의 정량적 분석)

  • Choi, Suin;Jeon, Pureum;Huh, Yang Hoon;Lee, Jin-A
    • Analytical Science and Technology
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    • v.31 no.5
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    • pp.208-218
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    • 2018
  • Autophagy is a cellular process whereby cytosolic materials or organelles are taken up in a double-membrane vesicle structure known as an autophagosome and transported into a lysosome for degradation. Although autophagy has been studied at the genetic, cellular, or biochemical level, systematic ultrastructural quantitative analysis of autophagosomes during the autophagy process by using transmission electron microscopy (TEM) has not yet been reported. In this study, we performed ultrastructural analysis of autophagosomes in wild-type (WT) mouse embryonic fibroblasts (MEFs) and autophagy essential gene (atg5) knockout (KO) MEFs. First, we performed ultrastructural analysis of autophagosomes in WT MEFs compared to atg5 KO MEFs in basal autophagy or starvation-induced autophagy. Although we observed phagopore, early, late autophagosomes, or autolysosomes in WT MEFs, atg5 KO MEFs had immature autophagosomes that showed incomplete closure. Upon starvation, late autophagosomes accumulated in WT MEFs while the number of immature autophagosomes significantly increased in atg5 KO MEF indicating that atg5 plays an important role in the maturation of autophagosomes. Next, we examined autophagosomes in the cell model expressing polyQ-expanded N-terminal fragment of huntingtin. Our TEM analysis indicates that the number of late autophagosomes was significantly increased in the cells expressing the mutant huntingtin, indicating that improving the fusion of autophagosome with lysosome may be effective to enhance autophagy for the treatment of Huntington's disease. Taken together, the results of our study indicate that ultrastructural and quantitative analysis of autophagosomes using TEM can be applied to various human cellular disease models, and that they will provide an important insight for cellular pathogenesis of human diseases associated with autophagy.

Histological Change of Uterus Endometrium and Expression of the Eggshell-related Genes according to Hen Age (닭의 산란연령에 따른 자궁내막조직의 변화 및 난각 관련 유전자의 발현양상)

  • Park, Ji Ae;Cho, Eun Jung;Park, Jung Yeon;Sohn, Sea Hwan
    • Korean Journal of Poultry Science
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    • v.44 no.1
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    • pp.19-28
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    • 2017
  • The eggshell is an intricate and highly ordered structure composed of multiple layers and a calcified matrix. The eggshell is formed at the uterine segment of the chicken oviduct. In this study, histological changes in the uterine endometrium and the expression of the eggshell-related genes were investigated according to hen age. We analyzed the expression of eggshell protein-related genes, such as OCX-32, OCX-36, OC-17, OC-116, and eggshell-ion-related genes, such as CABL-1, SPP1, SCNN1G, ATP2A2, CA2, and CALM1. In chicken uterine endometrium, histological deformation, fibrosis, atrophy and elimination of micro-villi were found with increasing hen age. The concentration of blood-ion components did not significantly change with age. The amount of telomeric DNA in uterine endometrial cells decreased with increasing hen age. The expression of most of the eggshell-related genes changed significantly with increasing hen age. The expression of some ovo-proteins, which play a role in eggshell formation, increased with increasing hen age; however, there were no significant correlations among eggshell protein genes. Eggshell ion-related genes, such as ATP2A2, SCNN1G, CA2, and CALM1, were closely related to each other. The OCX-32 and OCX-36 genes were closely related to some of the eggshell ion genes. Eggshell protein-related genes, such as the OCX-32, OCX-36 genes and ion-related genes such as CALB-1, ATP2A2, SCNN1G, CA2, CALM1, affected eggshell formation, mutually or independently. This study shows that, uterine although endometrial cell damage occurs with increasing hen age, normal eggshells can be formed in old hens. This suggests that eggshell protein-and eggshell ion-related genes also control the homeostasis of eggshell formation.