• Title/Summary/Keyword: 유전체 분석

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Development of Lipase Hyper-producing Strain from Hybrids between Aspergillus niger and Penicillium notatum by Nuclear Transfer (핵전이에 의한 Aspergillus niger와 Penicillium notatum 잡종에서의 lipase 고생산 균주의 개발)

  • Yang, Young-Ki;Moon, Myeng-Nim;Lee, Yoon-Hee;Kang, Hee-Kyoung;Lee, Jung-Sup;Lim, Chae-Young;Kim, Jong-Se;Rhee, Young-Ha
    • The Korean Journal of Mycology
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    • v.25 no.2 s.81
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    • pp.143-151
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    • 1997
  • Interspecific hybrids between Aspergillus niger and Penicillium notatum (Tyr-), hyperlipolytic enzyme-producing fungi, were obtained by nuclear transfer technique. Optimal conditions for formation of intergeneric hybrids were investigated. Maximum production of protoplasts was obtained by 1% Novozyme 234 at $30^{\circ}C$ for 3 hrs and the most effective osmotic stabilizers for the isolation of protoplasts were 0.6 M KCl. Frequencies of hybrid formation by nuclear transfer were $3.8{\times}10^{-3}{\sim}1.3{\times}10^{-3}$. From the observation of genetic stability, conidial size, DNA content, and nuclear stain, it was suggested that their karyotypes are aneuploid. The hybrids showed $1.2{\sim}1.7$ fold higher lipase activities than parental strains. It was strongly supported by results of this study that nuclear transfer technique is much more efficient in the formation of intergeneric hybrids than protoplast fusion and is very useful for the improvement of strains.

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In Vitro Construction and Characterization of the Bacteriophage P4 Derivative, P4 sid71 cosP2, Containing the Bacteriophage P2 cos Region (박테리오파지 P2의 cos 지역을 함유하는 박테리오파지 P4 유도체인 P4 sid71 cosP2의 In vitro 조성과 정성 연구)

  • Kim, Kyoung-Jin
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.99-104
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    • 2013
  • Bacteriophage P2 sir mutants are inefficient helpers for their satellite bacteriophage P4. The term, "P2 sir-associated helper inefficiency" has been used to define this phenomenon and it has been suggested that the DNA sequence difference between the cos region of P2 and that of P4 is responsible. To test this hypothesis, P4 derivative phage, P4 sid71 cosP2, containing the cos region of P2 and sid71 allele was constructed through several in vitro DNA manipulation steps. Its burst size was determined using a one-step growth experiment. The results showed that the substitution of the cos region of P2 for the cos region of P4 in P4 sid71 cosP2 overcame "P2 sir-associated helper inefficiency". P4 sid71 cosP2 stock phages prepared with P2 wild type helper and P2 sir helper were analyzed using a CsCl buoyant equilibrium density gradient experiment. The results revealed that the phage particles containing three copies of the P4 genome were the predominant particles in both cases.

Evaluation of Drought Stress in Native Kimchi Cabbage from Korea (국내 재래종 배추 유전자원의 내건성 평가)

  • Si-Hong Kim;Soo-Yong Choi;Eui-Sub Lee;Young-Hyun Woo;Jae-Yun Heo
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.08a
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    • pp.69-69
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    • 2020
  • 식물체가 건조 스트레스를 받으면 각 기관 물질 생산의 변이, 분화 및 발달 억제를 통해서 식물의 생산활동을 현저히 저하시켜, 식물의 생장, 형태, 개체발생 및 대사생리에 영향을 미치는 것으로 연구되어 왔다. 최근 기후 온난화로 인해 온도가 점진적으로 상승하고 가뭄과 같은 이상기상이 빈번하게 발생함에 따라 많은 노지 작물의 농업 생산성이 약화되고 있다. 배추는 우리나라의 대표적인 식품 중 하나인 김치의 주재료로 연중 안정적인 생산과 공급이 필요시 되지만, 배추의 경우 건조 조건에서 엽육조직의 붕괴와 같은 생리장해가 발생되기 때문에 최근 발생되고 있는 이상 기후의 영향으로 안정적인 생산이 어려워지고 있는 실정이다. 하지만, 배추에서는 이상 기후에 대응이 가능한 내건성 품종의 육성을 위한 연구는 미흡한 실정이다. 따라서 본 연구에서는 내건성이 높은 배추 개발을 위한 기초소재를 찾는 것을 목적으로 농촌진흥청 유전자원센터에서 보존하고 있는 재래종 배추 30계통을 분양받아 강릉원주대학교 생명과학대학 내의 조직배양실을 이용하여 연구를 수행하였다. 배지는 Tissue Culture Square Dish(125×125×20mm)에 Agar를 녹여 40ml씩 분주하여 고체배지를 조성한 후, 건조 처리구 Polyrthylene glycol 6000(PEG) 0%(Control), 20%(Mild Stress), 30%(Severe Stress)를 설정하여 60ml씩 추가 분주하여 배양기 28℃에서 15시간 처리를 하였다. 분양받은 각 계통의 종자는 1% 차아염소산 나트륨으로 10분간 종자표면을 살균한 후, 5번 정도 멸균수로 헹군 후, 표면 살균한 재래종 종자를 고체배지 시험관에 6립씩 치상하였다. 식물체 생육은 각 처리구별 3반복으로 하였으며 주간 12시간 주기, 광도 2,400Lux, 온도 20℃의 조직배양실에서 치상 후 7일간 생육하였다. 치상 후 7일간 생육시킨 식물체를 채취하여 지상부생체중, 지하부 생체중, 뿌리 길이를 측정하였으며, 지상부와 지하부로 나누어 50℃에서 72시간 건조시킨 후, 건물 생산량을 조사하였다. 본 실험 결과 건조 처리(PEG-6000)는 배추의 생장을 저해하였지만, 생육 저해 정도는 계통간의 차이가 있는 것을 관찰할 수 있었으며 무처리구와 스트레스 처리구간의 생장량 변화 정도를 기반으로 군집분석을 수행한 결과 'IT110483'과 'IT104903' 계통이 실험에 공시된 계통들 중 상대적으로 강한 건조 내성을 가진 것을 확인할 수 있었다. 따라서, 재래종 배추 계통 중 일부는 건조내성이 강한 새로운 배추 품종을 육성하는데 있어 유용하게 이용될 수 있을 것으로 사료된다.

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Speciation Mode Reconstruction for Lepilemur six species (L. mustelinus, L. edwardsi, L. dorsalis, L. leucopus, L. ruficaudatus, L. septentrionalis) Based on the Lepilemur Karyotype Analysis (여우원숭이속(Lepilemuridae)의 핵형 분석을 통해 나타난 Lepilemur 6종(L. mustelinus, L. edwardsi, L. dorsalis, L. leucopus, L. ruficaudatus, L. septentrionalis)의 종 분화 양상)

  • 정기윤
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.5 no.2
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    • pp.141-149
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    • 2004
  • The aim of this study was to test the validity of the hypothesis that the karyotypes of four species of Lepilemuridae were formed spontaneously from their ancestral hybrid karyotype. Hypothetical ancestral haploid Karyotype of Lepilemuridae is composed of 18 autosomes and X chromosomes. Lepilemur mustelinus karyotype has four tandem fused chromosomes and one Robertsonian translocated chromosome pairs. Lepilemur septentrionalis septentrionalis karyotype has only two pairs of translocated chromosomes. We reconstruct and suggest ancestral karyotype of LMU(ancLMU) and LSS(ancLSS), from which all four studied species were derived. Hybrids of ancLMU and ancLSS were formed and produce differently fused equilibrated gametes via circular form arrangement during gametogenesis. Five unit of trivalent homologous chromosome pairs were engaged in a circular form to give new gamete corresponding to the karyotype of L. dorsalis, orientation of one unit of trivalent was inversed in the circle to gave new gamete corresponding to the karyotype of L. leucopus. Seven homologous chromosome pairs were engaged in circular form to give haploid karyotype of Lepilemur ruficaudatus. Only one homologous chromosome pair is dissociated and the other chromosome pairs rearranged in the circle to form haploid karyotype of Lepilemur edwardsi. The new gametes could be produced from these circular forms. When the new gamete fertilized with the same type of gamete, The new homozygote is produced as existing L. dorsalis, L. leucopus, L. edwardsi and L. ruficaudatus. These results support the theory that new species could be formed in hybrid population through activated chromosome fusion, chromosome rearrangement in circular from at zygotene stage and production of equilibrated gametes to form homozygote new species.

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Molecular Epidemiology of Human Astrovirus Infection in Hospitalized Children with Acute Gastroenteritis (급성 위장관염으로 입원한 소아에서 분자 유전학적 방법에 의한 Human Astrovirus 감염의 유병률)

  • Chung, Ju-Young;Huh, Kyung;Kim, Sang Woo;Shin, Bo Mun;Han, Tae Hee;Lee, Jae In;Song, Mi-Ok
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.9 no.2
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    • pp.139-146
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    • 2006
  • Purpose: Human astrovirus (HAstV) is known to be an important etiologic agent of acute gastroenteritis in infants worldwide. However, the prevalence of HAstV infection varies according to geographic region and patient age. The purpose of our study was to investigate the incidence of HAstV infection among hospitalized children at a tertiary hospital in Seoul. Methods: Fecal samples were collected from hospitalized children up to five years of age with acute gastroenteritis. A total of 812 fecal samples were collected from hospitalized children with acute gastroenteritis between February 2004 and January 2005. Fecal specimens were screened for rotavirus, enteric adenovirus and norovirus by enzyme immunoassay (EIA) or reverse transcriptase polymerase chain reaction (RT-PCR). HAstV positive samples were characterized by RT-PCR. Results: Rotavirus was detected in 16.9% (138/812), norovirus in 11.6% (94/812), and adenovirus in 4.0% (33/812) of the study population. HAstV was detected in 4.0% (33/812) samples by RT-PCR. The age distribution of HAstV positive patients was as follows: <12 month old, 82.0% (27/33); 1~2 years old, 6.0% (2/33); 2~5 years old, 12.0% (4/33). The seasonal distribution of HAstV positive samples was as follows; April (3), May (5), June (4), August (12), September (4), October (2), November (2), and December (1). The peak rate of HAstV infection was observed during the summer season, 2004. A mixed infection of viral agents was confirmed in 2.7% (22 /812) of the study population, most commonly with rotavirus and norovirus, and with rotavirus and HAstV. Genotype 1 was the predominant type (91%, 20/22) and genotype 8 was detected in two cases. Conclusion: The prevalence of HAstV infection was 4.0% in hospitalized children with acute gastroenteritis, and was especially high in infants. HAstV can be considered as an important etiologic agent of gastroenteritis in children.

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Characterization of Exolytic GH50A β-Agarase and GH117A α-NABH Involved in Agarose Saccharification of Cellvibrio sp. KY-GH-1 and Possible Application to Mass Production of NA2 and L-AHG (Cellvibrio sp. KY-GH-1의 아가로오스 당화 관련 엑소형 GH50A β-아가레이즈와 GH117A α-NABH의 특성 및 NA2와 L-AHG 양산에의 적용 가능성)

  • Jang, Won Young;Lee, Hee Kyoung;Kim, Young Ho
    • Journal of Life Science
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    • v.31 no.3
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    • pp.356-365
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    • 2021
  • Recently, we sequenced the entire genome of a freshwater agar-degrading bacterium Cellvibrio sp. KY-GH-1 (KCTC13629BP) to explore genetic information encoding agarases that hydrolyze agarose into monomers 3,6-anhydro-L-galactose (L-AHG) and D-galactose. The KY-GH-1 strain appeared to possess nine β-agarase genes and two α-neoagarobiose hydrolase (α-NABH) genes in a 77-kb agarase gene cluster. Based on these genetic information, the KY-GH-1 strain-caused agarose degradation into L-AHG and D-galactose was predicted to be initiated by both endolytic GH16 and GH86 β-agarases to generate NAOS (NA4/NA6/NA8), and further processed by exolytic GH50 β-agarases to generate NA2, and then terminated by GH117 α-NABHs which degrade NA2 into L-AHG and D-galactose. More recently, by employing E. coli expression system with pET-30a vector we obtained three recombinant His-tagged GH50 family β-agarases (GH50A, GH50B, and GH50C) derived from Cellvibrio sp. KY-GH-1 to compare their enzymatic properties. GH50A β-agarase turned out to have the highest exolytic β-agarase activity among the three GH50 isozymes, catalyzing efficient NA2 production from the substrate (agarose, NAOS or AOS). Additionally, we determined that GH117A α-NABH, but not GH117B α-NABH, could potently degrade NA2 into L-AHG and D-galactose. Sequentially, we examined the enzymatic characteristics of GH50A β-agarase and GH117A α-NABH, and assessed their efficiency for NA2 production from agarose and for production of L-AHG and D-galactose from NA2, respectively. In this review, we describe the benefits of recombinant GH50A β-agarase and GH117A α-NABH originated from Cellvibrio sp. KY-GH-1, which may be useful for the enzymatic hydrolysis of agarose for mass production of L-AHG and D-galactose.

Changes of Protein Profiles in Cheonggukjang during the Fermentation Period (전통 청국장의 발효 기간 동안 변화하는 수용성 단백질 개요)

  • Santos, Ilyn;Sohn, Il-Young;Choi, Hyun-Soo;Park, Sun-Min;Ryu, Sung-Hee;Kwon, Dae-Young;Park, Cheon-Seok;Kim, Jeong-Hwan;Kim, Jong-Sang;Lim, Jin-Kyu
    • Korean Journal of Food Science and Technology
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    • v.39 no.4
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    • pp.438-446
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    • 2007
  • The fermented soybean product, cheonggukjang, is favored by many people, partly due to its bio-functional ingredients. Since the fermentation process of cheonggukjang is mediated by enzymes, including proteases, produced by microbes, analysis of the proteome profile changes in cheonggukjang during fermentation would provide us with valuable information for fermentation optimization, as well as a better understanding of the formation mechanisms of the bio-functional substances. The soluble proteins from cheonggukjang were prepared by a phenol/chloroform extraction method, in order to remove interfering molecules for high resolution 2-D gel analysis. Proteomic analysis of the cheonggukjang different fermentation periods suggested that most of the soluble soy proteins were degraded into smaller forms within 20hr, and many microbial proteins, such as mucilage proteins, dominated the soluble protein fraction. The proteomic profile of cheonggukjang was very different from natto, in terms of the 2-D gel protein profile. Among the separated protein spots on the 2-D gels, 50 proteins from each gel were analyzed by MALDI-TOF MS and PMF for protein identification. Due to database limitations with regard to soy proteins and microbial proteins, identification of the changed proteins during fermentation was restricted to 9 proteins for cheonggukjang and 15 for natto. From de novo sequencing of the proteins by a tandem MS/MS, as well as by database searches using BLASTP, a limited number of proteins were identified with low reliability. However, the 2-D gel analysis of proteins, including protein preparation methods, remains a valuable tool to analyze complex mixtures of proteins entirely. Also, for intensive mass spectrometric analysis, it is also advisable to focus on a few of the interestingly changed proteins in cheonggukjang.

Multiple accelerated degradation test and failure analysis for $Ni-BaTiO_3$ MLCCs ($Ni-BaTiO_3$ MLCCs에 대한 복합 가속 열화 시험 및 고장 분석)

  • Kim, Jung-Woo;Kim, Jin-Seong;Lee, Hee-Soo;Kang, Do-Won;Kim, Jeong-Wook
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.19 no.2
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    • pp.102-105
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    • 2009
  • The accelerated life time test of the MLCCs with different $BaTiO_3$ particle sizes were conducted at $150^{\circ}C$, 75 V condition and the effect of $BaTiO_3$ particle size on the breakdown voltage and degradation characteristics of MLCCs was investigated. The MLCCs were prepared by using the $BaTiO_3$ particles having the size of $0.525{\mu}m$, $0.555{\mu}m$, $0.580{\mu}m$ and Ni-electrode, respectively. The MLCCs which have the particle size of $0.525{\mu}m$, $0.555{\mu}m$, and $0.580{\mu}m$, respectively were confirmed to meet the standard requirements of X5R(change capacitance within ${\pm}15%$ at $-55{\sim}85^{\circ}C$) by TCC(Temperature Coefficient of Capacitance). The effect of the $BaTiO_3$ particle size on the insulation resistance behavior of MLCCs was confirmed by BDV(Breakdown Voltage) measurements and the cause and degree of degradation of MLCCs were characterized by XPS analysis after the accelerated life test. The MLCCs with $0.525{\mu}m-BaTiO_3$ showed better insulation resistance and BDV characteristics compare to other MLCCs and XPS analysis revealed that the MLCCs degradation is caused by the NiO peak and $BaTiO_3$ peak decrease.

Molecular epidemiological study of norovirus gastroenteritis outbreaks in Gyeonggi-Do from 2014 to 2015 (2014-2015년 경기지역 노로바이러스성 식중독의 분자역학적 특성분석)

  • Nam, Soo-Jung;Park, Po-Hyun;Bang, Sun-Jae;Huh, Jeong-Weon;Yun, Hee-Jeong;Park, Kwang-Hee;Yoon, Mi-Hye
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.24-30
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    • 2018
  • Norovirus infection is a leading cause of nonbacterial gastroenteritis outbreaks. New variants of GII.4 have emerged approximately every 2~3 years and have caused norovirus gastroenteritis pandemics globally. In this study, analysis and molecular genetic characteristics of the norovirus gastroenteritis outbreaks 2,917 samples in Gyeonggi-Do from 2014 to 2015. As a result, 247 samples out of 2,917 samples are positive for norovirus. Norovirus molecular genetic characteristics of the GI 8 types (GI-1, 2, 3, 4, 5, 6, 12, 14), GII 10 types (GII - 2, 3, 4, 5, 6, 11, 12, 14, 16, 17). Genome sequences of isolated noroviruses were similar to those of new GII.17 Kawasaki 2014 variants with 96.6 identity, suggesting that these viruses were imported from overseas. 44% of virus incidence was originated from school meal service. Therefore, a continuous monitoring and school sanitation should be required for preventing a massive virus outbreak.

Genetic Character and Essential Oil Components of Domestic Agastache rugosa (Fisch. & Mey.)Kuntze (국내 재생 배초향(Agastache rugosa (Fisch. & Mey.)Kuntze)의 형질특성 및 정유성분)

  • Ki Yeon Lee;A-Reum Park;Jae Hee Lee;Hee Yeon Kim;Sung Jin Choi;Jin Gwan Ham
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.12a
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    • pp.57-57
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    • 2020
  • 본 연구는 국내 자생 방향성 식물의 자원발굴과 증식, DB 구축을 위하여 수행되었다. 배초향은 꿀풀과에 속하는 다년생 식물로 대표적인 한국의 방향식물이다. 한의학에서는 배초향의 지상부를 곽향이라 하여 음식으로 인한 체증, 오한, 복통 등을 치료하는 한약재로 사용되며 그 밖의 관상용, 밀원용, 식품용, 향신료 등 다양한 산업제품의 원료로 사용되고 있다. 배초향의 유전형질 특성 및 정유성분 조사를 위하여 국립농업유전자원센터로부터 배초향 종자(30자원)를 분양받아 발아시킨 후 시험연구포장에 증식하였다. 증식된 배초향을 대상으로 개화기, 식물 및 꽃의 색, 잎의 모양 등 개체별 형질특성을 조사하였으며 일정시기에 일시 수확하여 정유추출 후 GC/MS 분석을 통하여 정유의 화학성분을 분석하였다. 배초향의 개화시기는 6월 10일 경~7월 29일 경이었고 초장은 약 80cm~186cm이었다. 잎의 모양은 30자원 중 광난형 12자원, 피침형 10자원, 난형 8자원의 분포였고 식물체의 색은 green과 red-purple 계열이 각각 22자원과 8자원이었다. 배초향 꽃의 색깔은 RHS Colour chart 참조하여 149P 10P 8/4, 129B 10P 7/8, 37V 2.5RP 4/12의 색으로 분류하여 조사하였다. 129B 10P 7/8에 해당하는 자원이 18자원으로 가장 많은 분포를 차지하였다. 배초향의 정유성분 분석 결과, 정유수율은 0.11%~0.44%이었고 정유성분은 estragole, methyl eugenol, menthone 등이었다. 배초향의 주성분으로 알려진 estragole은 30자원 중 13자원의 주요정유성분인 것으로 나타났다.

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