• 제목/요약/키워드: DNA amplification

검색결과 888건 처리시간 0.033초

Evaluation of Yeast Diversity During Wine Fermentations with Direct Inoculation and pied de cuve Method at an Industrial Scale

  • Li, Erhu;Liu, Chuanhe;Liu, Yanlin
    • Journal of Microbiology and Biotechnology
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    • 제22권7호
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    • pp.960-966
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    • 2012
  • The diversity and composition of yeast populations may greatly impact wine quality. This study investigated the yeast microbiota in two different types of wine fermentations: direct inoculation of a commercial starter versus pied de cuve method at an industrial scale. The pied de cuve fermentation entailed growth of the commercial inoculum used in the direct inoculation fermentation for further inoculation of additional fermentations. Yeast isolates were collected from different stages of wine fermentation and identified to the species level using Wallersterin Laboratory nutrient (WLN) agar followed by analysis of the 26S rDNA D1/D2 domain. Genetic characteristics of the Saccharomyces cerevisiae strains were assessed by a rapid PCR-based method, relying on the amplification of interdelta sequences. A total of 412 yeast colonies were obtained from all fermentations and eight different WL morphotypes were observed. Non-Saccharomyces yeast mainly appeared in the grape must and at the early stages of wine fermentation. S. cerevisiae was the dominant yeast species using both fermentation techniques. Seven distinguishing interdelta sequence patterns were found among S. cerevisiae strains, and the inoculated commercial starter, AWRI 796, dominated all stages in both direct inoculation and pied de cuve fermentations. This study revealed that S. cerevisiae was the dominant species and an inoculated starter could dominate fermentations with the pied de cuve method under controlled conditions.

가지의 Ralstonia solanacearum에 의한 풋마름병 발생과 생리형의 분화 (Occurrence and Biovar Classification of Bacterial Wilt Caused by Ralstonia solanacearum in Eggplant (Solanum melongena))

  • 임양숙;이문중;정종도;류영현;김병수
    • 식물병연구
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    • 제14권1호
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    • pp.10-14
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    • 2008
  • Ralstonia solanacearum에 의한 풋마름병은 세계적으로 치명적인 병해중의 하나이며 우리나라에서는 가지, 토마토, 감자, 고추 등 가지과 작물에 발병이 보고되어 있다. 2005년${\sim}$2006년 2년 동안 가지 주재배단지의 풋마름병 발생 포장에서 48개의 균주를 수집하여 분리 배양하였으며 R. solanacearum의 특이적인 flipcF/flipcR primer를 이용하여 PCR을 실시한 결과 470-bp의 풋마름병 특이적 밴드가 관찰되어 R. solanacearum으로 동정하였다. 분리 동정된 15균주로 각 기주별 병원성을 검정하였을 때 가지, 토마토에 병원성을 나타내었으나 고추는 저항성을 나타내었다. 가지 풋마름병원균의 biovar분화는 biovar 3, 4로 동정되었으며 특히 biovar 3는 가지 풋마름병 저항성 대목에 다소 강한 병원성을 나타내어 저항성대목의 육종이 필요한 것으로 판단된다.

SSR 마커를 이용한 산양삼의 유전적 다양성 분석 (Genetic Diversity Analysis of Wood-cultivated Ginseng using Simple Sequence Repeat Markers)

  • 길진수;엄유리;변재경;정종욱;이이;정찬문
    • 한국약용작물학회지
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    • 제25권6호
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    • pp.389-396
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    • 2017
  • Background: Panax ginseng C. A. Meyer is wood-cultivated ginseng (WCG) in Korea which depends on an artificial forest growth method. To produce this type of ginseng, various P. ginseng cultivars can be used. To obtain a WCG similar to wild ginseng (WG), this method is usually performed in a mountain using seeds or seedlings of cultivated ginseng (CG) and WG. Recently, the WCG industry is suffering a problem in that Panax notoginseng (Burk.) F. H. Chen or Panax quinquefolium L. are being sold as WCG Korean market; These morphological similarities have created confusion among customers. Methods and Results: WCG samples were collected from five areas in Korea. After polymerase chain reaction (PCR) amplification using the primer pair labeled with fluorescence dye (FAM, NED, PET, or VIC), fragment analysis were performed. PCR products were separated by capillary electrophoresis with an ABI 3730 DNA analyzer. From the results, WCG cultivated in Korea showed very diverse genetic background. Conclusions: In this study, we tried to develop a method to discriminate between WCG, P. notoginseng or P. quinquefolium using simple sequence repeat (SSR) markers. Furthermore, we analyzed the genetic diversity of WCG collected from five cultivation areas in Korea.

16S rRNA 유전자 계통분석에 의한 한강수계의 세균 다양성 (Bacterial Diversity of the Han River as Determined by 16S rRNA Gene Analysis)

  • 한석균;이일규;안태영
    • 미생물학회지
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    • 제34권4호
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    • pp.194-199
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    • 1998
  • 한강의 본류와 만나는 탄천과 중랑천에서 16S rDAN를 증폭하고 부분적인 염기서열 분석을 통하여 한강의 세균 다양성을 결정하였다. 총 27개의 클론을 분리하였으며 RFLP를 이용하여 7개의 group으로 나누었다. 탄천의 15개 클론은 4개의 group으로 나뉘어졌으며 가장 많은 클론을 포함하는 group(HT-1 클론)은 class Proteobacteria의 ${\delta}$-subdivision에 속하는 Acrobacter cryaerophilius와 높은 유사도를 보였으며, 다른 두 group(HT-6과 HT-9 클론)은 모두 clas Cytophagales에 속하였다. 중랑천의 12개의 클론은 3개의 group으로 나뉘어졌으며 가장 많은 클론을 보이는 group(HJ-1 클론)은 class Proteobacteria의 ${\alpha}$-subdivision에 속하는 Sphingomonas sp. 와 높은 유사도를 나타내었다. 전체적으로는 Proteobateria(alpha, beta and delta subdivision), Cytophagales와 Actinomycetales가 검출되었다.

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오징어과의 Kinesin Superfamily Proteins (KIFs)의 유전자분석 및 계통분석 (Sequences and Phylogenic Analysis of Squid New Kinesin Superfamily Proteins (KIFs))

  • 김상진;석대현
    • 생명과학회지
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    • 제22권3호
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    • pp.293-297
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    • 2012
  • 분자 운동 단백질은 신경세포 내의 세포체에서 특정 목적지까지 소포를 이동시키는데 관여한다. 오징어의 거대 축삭은 간단한 제거조작으로 축삭을 분리 가능하기 때문에 신경세포내 물질이동기전 연구의 좋은 모텔로 활용 가능하다. 이전연구에서 오징어 거대축삭의 소포들은 미세소관을 따라 이동하는 키네신 항체에 의하여 운반됨이 확인되었다. 본 연구는 오징어 뇌에 존재하는 키네신들을 크로닝하고, 분리된 유전자의 분석을 행하기 위하여 키네신 운동 도메인에서 잘 보존된 아미노산 배열에 해당되는 영역에 DNA primer을 이용하여 새로운 6종류의 키네신을 분리하였다. 오징어의 키네신들과 생쥐의 키네신들의 motor 영역의 아미노산분석에서 보존된 영역이 존재하며, Maximum Parsimony (MP) 방법, Neighbor-Joining (NJ) 방법, Minimum Evolution (ME) 방법, 그리고 Maximum likelihood (ML) 방법을 기초로 한 계통분석에서 생쥐의 키네신과 높은 상동성을 나타내었으며, 또한 계통수에서도 높은 상관관계가 확인되었다.

반복적인 의식변화와 운동실조를 주소로 진단된 지발형 Ornithine Transcarbamylase Deficiency 1례 (A Case of Late Onset Ornithine Transcarbamylase Deficiency in a 6 Year-old Girl Who Showed Recurrent Episodic Mental Changes and Ataxia)

  • 정권;김은영;김경심;김용욱;유한욱
    • 대한유전성대사질환학회지
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    • 제3권1호
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    • pp.32-37
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    • 2003
  • 저자들은 반복적이며 간헐적인 기면, 정신착란과 운동실조를 주소로 내원한 6세 여아에서, 고암모니아혈증과 혈장 glutamine, 요 orotic acid의 증가를 보여 지발형 OTC 결핍증으로 진단하고, 분자유전학적 검사상 exon 6에서 221번째 아미노산 lysine에 해당하는 염기 AAG가 AAT(asparagine)로 치환된 돌연변이를 이형접합자(heterozygote)로 보인하였던 1례를 경험하였기에 보고하는 바이다.

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인체 뇌종양조직에서 텔로머레이즈의 발현과 세포사멸 (Expression of Telomerase Activity and Apoptosis in Human Brain Tumors)

  • 김충현;정진환;백광흠;김재민;고용;오석전
    • Journal of Korean Neurosurgical Society
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    • 제30권2호
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    • pp.137-143
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    • 2001
  • Objective : Telomerase, a ribonucleoprotein adds telomere repeats to the ends of telomeres to compensate for the progressive loss. A favorable prognosis associated with low or no telomerase activity in some tumors, and cells transfected with antisense human telomerase lost telomeric repeats and die. We studied about the relationship between telomerase activity and apoptosis in the human brain tumors. Material and Methods : Between July 1998 and December 1999, 62 patients with brain tumors underwent surgery and their surgical specimens were obtained. Telomerase activity was investigated by telomeric repeats amplification protocol(TRAP) assay. Apoptosis was also evaluated by DNA fragmentation analysis. Differences and correlation in data were analyzed using Mann-Whitney test and Wilcoxon-signed rank test. Results : Expression rate of telomerase activity and apoptosis were 80% and 30% in malignant gliomas, 33% and 0% in low grade gliomas, 63% and 38% in meningiomas, 67% and 33% in pituitary adenomas, 33% and 33% in metastatic tumors, 67% and 17% in acoustic neurinomas, 100% and 100% in pineoblastomas, 100% and 0% in the hemangioblastoma, respectively. There was no significant difference of telomerase activity and apoptosis between histological types. But a significant difference was noted in the expression of telomerase activity between malignant gliomas and low grade gliomas(p = 0.022). Brain tumors with telomerase activity expressed the lower rate of apoptosis. A significant correlation was also found between telomerase activity and absence of apoptosis in the human brain tumors(p = 0.005). Conclusions : Our data suggests that telomerase may protect from apoptosis of the human brain tumors and also may play an important role in the biological malignancy of the gliomas.

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Ethidium monoazide (EMA) - PCR 법을 이용한 비배양성 생존 상태(VBNC)의 Edwardsiella tarda 검출 (Differentiations between the viable but nonculturable (VBNC) or dead state of Edwardsiella tarda by ethidium monoazide (EMA) treatment-PCR)

  • 강남이;김은희
    • 한국어병학회지
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    • 제31권2호
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    • pp.93-99
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    • 2018
  • Edwardsiella tarda predominantly causes edwardsiellosis in fish at high temperature, but is rarely isolated from water when water temperature is low. However, E. tarda is viable but nonculturable (VBNC) in low water temperature, but it can be revived when water temperature rises and cause disease to fish. Therefore, in order to prevent disease, it is very important to identify pathogens that are in the VBNC state in environmental water. In this study, E. tarda cells in the VBNC state were detected by the ethidium monoazide (EMA)-PCR method using the low-temperature oligotrophic sea water microcosm obtained by inoculation of E. tarda at a concentration of $10^8CFU/ml$. In order to distinguish between live and dead bacteria in E. tarda, each sample was treated with EMA at different concentrations, photoactivated with a 500 W halogen lamp, and PCR was performed with E. tarda specific primer. At the concentration of $10^7CFU/ml$ bacterium, DNA amplification was observed only in the live cells when treated with $60{\mu}g/ml$ of EMA, and smaller amounts of live cells could be distinguished from dead cells by adjusting the EMA concentration. In addition, the VBNC cells of E. tarda in the oligotrophic low temperature seawater microcosm were estimated to be in the range of $10^4{\sim}10^5CFU/ml$ by EMA-PCR. Therefore, it is possible to detect VBNC cells that will act as potential pathogens in environmental water using EMA-PCR method, and quantitative confirmation using concentration change is also possible.

Genetic diversity analysis of high yielding rice (Oryza sativa) varieties cultivated in Bangladesh

  • Epe, Isma Akter;Bir, Md. Shahidul Haque;Choudhury, Abul Kashem;Khatun, Asma;Aktar, Most Mohshina;Arefin, Md. Shamsul;Islam, Mohammed Aminul;Park, Kee Woong
    • 농업과학연구
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    • 제48권2호
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    • pp.283-297
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    • 2021
  • Investigation of genetic diversity and molecular characterization in high yielding rice varieties is important for their identification. The experiment was conducted during 2016 - 2017 to analyse the genetic diversity of fifteen high yielding rice varieties in Bangladesh by using random amplification of polymorphic DNA (RAPD) markers. Polymorphism was revealed in 12 RAPD primers out of 30, whereas no other reaction was detected on the remaining 18 primers. The 40 out of 45 bands (88.89%) polymorphics were produced by the primers and ranged from 50 to 100%. The maximum number of polymorphic bands was produced by the primer OPB-18 whereas the lowest number of polymorphic bands belonged to OPC-12. The genetic similarity coefficients were determined with the RAPD data, which ranged from 0.47 to 0.94. The unweighted paired group of arithmetic means (UPGMA) dendrogram presented the studied rice varieties into two major clusters. Moreover, the value of Nei's genetic diversity is 0.26 and the Shanon information index is 0.41. The study produced distinct positions, suggesting that the genotypes were different from each other. The results indicated that these markers could be efficient for comparing the genetic relationships, patterns of variation, and measurement of genetic distance among rice varieties. Considering all of these results, RAPD analysis is found to be an effective tool for estimating the genetic diversity of different rice varieties. The outcomes of this research may contribute to the germplasm data of rice accessions and a future breeding program of rice genotypes.

봉변에서 특이 유전자 검출법에 의한 봉군 내 꿀벌가시응애류 (Tropilaelaps)의 정량적 검출 (Quantitative Detection of Tropilaelaps in Hive by Specific Gene Detection from Hive Debris)

  • 김병희;김소민;김문정;김정민;;김선미;윤병수
    • 한국양봉학회지
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    • 제34권1호
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    • pp.27-37
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    • 2019
  • Rapid detection of Tropilaelaps, an external parasite of honeybees that lead to malformation of honeybee or colony collapse disorder, is becoming important. But it is very difficult to find with the naked eye of Tropilaelaps. In this study, we have developed a method to detect the specific gene of Tropilaelaps from the hive debris and to know the number of Tropilaelaps in the hive through Tropilaelaps-specific quantitative detection. Tropilaelaps-specific gene amplified in DNA extracted from hive debris by consecutive PCR (1st detection, 2nd nested PCR). It could detect 101 molecules level of Tropilaelaps-specific gene and confirm the amplification of the Tropilaelaps-specific gene. It was possible to accurately quantify the number of Tropilaelaps from the hive debris sample, which is difficult to discriminate the presence of Tropilaelaps visually, through Tropilaelaps-specific detection. Under the microscope, Tropilaelaps was collected and quantitative detection of Tropilaelaps-specific genes was performed. It was possible to quantify the number of Tropilaelaps present in the hive through the molecules of the quantified Tropilaelaps-specific genes. We suggest that hive debris can represent as a micro-environment to hive and show that it can be a simpler and more accurate sample than using a parasitic host honeybee. We expect that hive debris should facilitate the monitoring of Tropilaelaps in hive.