• 제목/요약/키워드: Reverse Transcription

검색결과 1,361건 처리시간 0.036초

A genetic approach to comprehend the complex and dynamic event of floral development: a review

  • Jatindra Nath Mohanty;Swayamprabha Sahoo;Puspanjali Mishra
    • Genomics & Informatics
    • /
    • 제20권4호
    • /
    • pp.40.1-40.8
    • /
    • 2022
  • The concepts of phylogeny and floral genetics play a crucial role in understanding the origin and diversification of flowers in angiosperms. Angiosperms evolved a great diversity of ways to display their flowers for reproductive success with variations in floral color, size, shape, scent, arrangements, and flowering time. The various innovations in floral forms and the aggregation of flowers into different kinds of inflorescences have driven new ecological adaptations, speciation, and angiosperm diversification. Evolutionary developmental biology seeks to uncover the developmental and genetic basis underlying morphological diversification. Advances in the developmental genetics of floral display have provided a foundation for insights into the genetic basis of floral and inflorescence evolution. A number of regulatory genes controlling floral and inflorescence development have been identified in model plants such as Arabidopsis thaliana and Antirrhinum majus using forward genetics, and conserved functions of many of these genes across diverse non-model species have been revealed by reverse genetics. Transcription factors are vital elements in systems that play crucial roles in linked gene expression in the evolution and development of flowers. Therefore, we review the sex-linked genes, mostly transcription factors, associated with the complex and dynamic event of floral development and briefly discuss the sex-linked genes that have been characterized through next-generation sequencing.

Silencing of the Target of Rapamycin Complex Genes Stimulates Tomato Fruit Ripening

  • Choi, Ilyeong;Ahn, Chang Sook;Lee, Du-Hwa;Baek, Seung-A;Jung, Jung Won;Kim, Jae Kwang;Lee, Ho-Seok;Pai, Hyun-Sook
    • Molecules and Cells
    • /
    • 제45권9호
    • /
    • pp.660-672
    • /
    • 2022
  • The target of rapamycin complex (TORC) plays a key role in plant cell growth and survival by regulating the gene expression and metabolism according to environmental information. TORC activates transcription, mRNA translation, and anabolic processes under favorable conditions, thereby promoting plant growth and development. Tomato fruit ripening is a complex developmental process promoted by ethylene and specific transcription factors. TORC is known to modulate leaf senescence in tomato. In this study, we investigated the function of TORC in tomato fruit ripening using virus-induced gene silencing (VIGS) of the TORC genes, TOR, lethal with SEC13 protein 8 (LST8), and regulatory-associated protein of TOR (RAPTOR). Quantitative reverse transcription-polymerase chain reaction showed that the expression levels of tomato TORC genes were the highest in the orange stage during fruit development in Micro-Tom tomato. VIGS of these TORC genes using stage 2 tomato accelerated fruit ripening with premature orange/red coloring and decreased fruit growth, when control tobacco rattle virus 2 (TRV2)-myc fruits reached the mature green stage. TORC-deficient fruits showed early accumulation of carotenoid lycopene and reduced cellulose deposition in pericarp cell walls. The early ripening fruits had higher levels of transcripts related to fruit ripening transcription factors, ethylene biosynthesis, carotenoid synthesis, and cell wall modification. Finally, the early ripening phenotype in Micro-Tom tomato was reproduced in the commercial cultivar Moneymaker tomato by VIGS of the TORC genes. Collectively, these results demonstrate that TORC plays an important role in tomato fruit ripening by modulating the transcription of various ripening-related genes.

엔테로바이러스 검출을 위한 real-time nucleic acid sequence-based amplification (NASBA), reverse transcription-PCR (RT-PCR) 및 바이러스 배양법의 비교 (Comparison of the Real-Time Nucleic Acid Sequence-Based Amplification (NASBA) Assay, Reverse Transcription-PCR (RT-PCR) and Virus Isolation for the Detection of Enterovirus RNA.)

  • 나영란;조현철;이영숙;빈재훈;최홍식;민상기
    • 생명과학회지
    • /
    • 제18권3호
    • /
    • pp.374-380
    • /
    • 2008
  • 본 연구는 무균성수막염 의심환자의 다양한 검체로부터 enterovirus의 진단을 위하여 real-time NASBA, 2 step RT-PCR 시험과 세포배양 시험을 각각 실시하여 각 시험법의 검출율, 특이도, 사용자 편리성, 시험소요 시간, 교차오염의 가능성 등을 비교 검토하였다. 비교시험 결과 전체 292건의 검체로부터 real-time NASBA에서 145건, 세포배양에서 101건, 2 step RT-PCR에서 86건이 양성으로 나타나 real-time NASBA가 가장 검출율이 높은 시험법임을 알 수 있었다. Enterovirus 외의 무균성수막염 원인바이러스에 대한 특이도 비교 시험결과 2 step RT-PCR 시험에서 rhinovirus 10건 중 1건이 위양성 반응을 나타내어 다른 시험법에 비해 특이도가 떨어지는 것으로 나타났다. Real-time NASBA는 하나의 튜브에서 증폭과 검출이 동시에 일어나 다른 시험과 비교하여 교차오염의 가능성이 낮으며 또한 시험 소요시간이 5시간 정도로 세포배양(5-14일 소요) 및 2 step RT-PCR(9시간소요) 에 비하여 신속하게 진단할 수 있어 일선병원이나 실험실에서 enterovirus를 검출을 위하여 적용할 수 있을 것으로 사료된다.

RAW 264.7을 이용한 정금나무 열매(Vaccinum oldhami fruit)의 항염증 효과 (Anti-inflammatory Activities Verification of Vaccinum oldhami Fruit Ethanol Extracts on RAW 264.7)

  • 이진영;주다혜;유단희;채정우
    • 생명과학회지
    • /
    • 제27권4호
    • /
    • pp.417-422
    • /
    • 2017
  • 본 연구는 천연물 소재인 정금나무 열매(Vaccinum oldhami fruit) 에탄올 추출물을 이용하여 항염증 효과에 대하여 검증하였다. 대식세포인 RAW 264.7 세포를 이용하여 정금나무 열매 추출물로 처리했을 때 세포 생존율을 확인한 결과 $500{\mu}g/ml$ 농도에서 118%를 나타내었다. 대식세포를 이용하여 정금나무 열매 추출물의 농도가 증가함에 따라서 변화하는 NO 생성량을 측정한 결과 최고농도 $1,000{\mu}g/ml$에서 47.3%로 감소한 것을 확인할 수 있었다. 정금나무 열매 에탄올 추출물을 Western blot을 이용하여 단백질 발현을 측정한 결과 처리한 세포군에서 농도가 증가함에 따라 iNOS와 COX-2의 단백질 발현양이 감소하여 최고 농도인 $500{\mu}g/ml$에서 각각 36.13%, 29.61%의 발현 억제를 보여주었다. Reverse-transcription PCR을 통하여 mRNA 발현양을 측정한 결과 iNOS와 COX-2의 mRNA 발현양이 감소하여 $500{\mu}g/ml$ 농도에서 62.25%, 90.07%로 내었다. 이러한 결과로 보아 정금나무 열매 에탄올 추출물의 항염증에 관한 그 효능을 확인할 수 있었고, 따라서 정금나무 열매 추출물을 이용한 천연 항염증 화장품 소재개발 이용 가능성을 넓힐 수 있을 것으로 사료된다.

Slow Bee Paralysis Virus (SBPV) 신속 검출을 위한 초고속 역전사 중합효소 연쇄반응법의 개발 (Development of Ultra-Rapid Reverse-Transcription PCR for the Rapid Detection against Slow Bee Paralysis Virus (SBPV))

  • 김소민;임수진;김정민;임윤규;윤병수
    • 한국양봉학회지
    • /
    • 제32권3호
    • /
    • pp.171-180
    • /
    • 2017
  • Slow Bee Paralysis Virus (SBPV)는 꿀벌 그리고 뒤영벌의 병원성 바이러스로써, 벌의 앞발을 마비시킴으로 폐사를 야기한다. 본 연구는, 뒤영벌에서 빠르게 SBPV 검출하고자 SBPV 특이 초고속 PCR법을 개발한 것을 보고하는 것이다. SBPV 특이 초고속 PCR은 제반 조건을 최적화한 후, $1.0{\times}10^8$ 분자의 SBPV 특이 유전자의 존재를 3분 35초 만에 인식할 수 있었으며, 최소 $1.0{\times}10^1$ 분자의 DNA까지 정량적으로 측정할 수 있었다. 한편, 국내외에서 생산된 뒤영벌을 시료로 SBPV 특이 초고속 역전사 PCR을 시행하였으며, 수입된 뒤영벌과 국내생산 뒤영벌 1종에서 SBPV가 존재함을 보일 수 있었다. 국내 처음으로 뒤영벌에서의 SBPV의 존재를 확인한 것이다. SBPV 특이 유전자 초고속 역전사 PCR법은 실험실 내에서 뿐만 아니라 현장에서 정량적 검출을 하기에 유용한 방법이 될 것이며 더 나아가 뒤영벌 수출입을 위한 검역 기준을 이용하여 집단적 발병 방지에 기여하기를 기대한다.

Expression of Enterotoxin Genes in Staphylococcus aureus Isolates Based on mRNA Analysis

  • Lee, Young-Duck;Moon, Bo-Youn;Park, Jong-Hyun;Chang, Hyo-Ihl;Kim, Wang-June
    • Journal of Microbiology and Biotechnology
    • /
    • 제17권3호
    • /
    • pp.461-467
    • /
    • 2007
  • Staphylococcus aureus strains are important foodborne pathogens that produce various toxins. To evaluate the risk of the enterotoxins, four S. aureus strains from kimbap and two clinical samples were isolated and identified, and their expression of the enterotoxin genes were analyzed using a reverse transcription real-time PCR. Various enterotoxin genes were detected, including sea, seg, seh, sei, sen, seo, and sem, where each isolate contained one or two. When the mRNA detection of the enterotoxin genes was analyzed using a reverse transcriptase PCR, various levels of expression were found depending on the species and enterotoxin gene. Therefore, it is reasonable to suggest that the poisoning risk of S. aureus can be effectively evaluated based on the gene expression at the mRNA level.

Development of Anti-viral Agents from Natural Sources

  • Hattori, Masao
    • Plant Resources
    • /
    • 제4권3호
    • /
    • pp.192-195
    • /
    • 2001
  • Human immunodeficiency virus (HIV), the causative agent of AIDS, still continues to spread rapidly in the world population, especially in Africa and Southeast Asia. At present, two kinds of therapeutic approaches are used for treatment of AIDS. One is to target HIV reverse transcriptase, which is responsible for the viral genome transcription. The other is to inhibit HIV pretense PR, which is essential for the processing of viral proteins. Drug combinations based on these approaches can reduce the blood virus to an undetectable level. However, a small amount of virus may lurk inside the immune cells in a dormant state. Another major obstacle of long-term treatment of the disease is remarkable mutation in HIV. Most of the clinical chemotherapeutic agents have one or more of these problems. High cost and harmful side-effects further reduced the desirability of these drugs. In the course our studies on development of anti-HIV agents from natural products, we investigated various crude drugs for their inhibitory activity against HIV-induced cytopathic effects (CPE) in culture cells, HIV-pretense (PR), HIV-reverse transcriptase (RT) including ribonuclease H (RNase H), and HIV integrase (INT). In the present paper, some inhibitory substances relating to the development of anti-HIV agents are reported.

  • PDF

풀망둑 난황전구단백질 유전자발현 추적기법 (Analysis of Vitellogenin Gene Expression in Synechogobius hastus (Gobiidae))

  • 계명찬
    • 환경생물
    • /
    • 제22권1호
    • /
    • pp.206-212
    • /
    • 2004
  • In an effort to develop the tools for monitoring the contamination of xenoestrogen in the aquatic environment of Korea, reverse transcription-polymerase chain reaction (RT-PCR) analysis of vitellogenin (VTG) mRNA expression were optimized in Synechogobius hastus. Based on the partial VTG cDNA sequence VTG mRNA level in livers from male fishes was analyzed by RT-PCR. As an internal control beta actin mRNA was amplified. 3 ${\mu}g$ of total RNA was reverse transcribed in 20 $\mu$l reaction using murine leukemia virus 〔MuLV〕 reverse transcriptase. Subsequent PCR using the 1 ${\mu}g$ of cDNA resulted in linear increase in PCR product of VTG in female liver cDNA from 10 to 30 cycles of amplification. On the contrary, in male, PCR product first detected at 28 cycles of amplification and linearly increased during 38 cycles of amplification, suggesting that male S. hastus expresses minute amount of VTG mRNA which is $2^{-18}$ equivalent of female. In conclusion, the optimized protocol of VTG mRNA expression in the liver of male S. hastus will be promising the environmental monitoring the xenoestrogen contamination in the western coast and estuaries in Korea.

역전사효소(逆轉寫酵素) 유전자(遺傳子)의 cloning 에 관(關)한 연구(硏究) (Cloning of Reverse Transcriptase Gene of Avian Sarcoma Virus)

  • 김용웅;김광식;서용택
    • Applied Biological Chemistry
    • /
    • 제31권3호
    • /
    • pp.219-225
    • /
    • 1988
  • Avian Sarcoma Virus의 plasmid DNA중(中)의 역 이사효소의 유전자(遺傳子)를 온도의존성(溫度依存性) 발현(發現) vector인 pPL-lambda에 cloning하여 온도(溫度)에 민감한 phage ${\lambda}$의 repressor인 cI857 gene을 갖고 있는 bacteriophage lysogen인 N4830에 transformation시켰다. transformant를 pL promoter의 발현(發現)을 억제(抑制)하는 저온(低溫)$(28^{\circ}C)$에서 배양(培養)시킨 뒤, 이 repressor를 억제(抑制)하여 transcription을 촉진(促進)하게 하는 고온(高溫)$(42^{\circ}C)$에서 배양(培養)시킨 다음 균체(菌體)를 회수(回收)하여 RNA를 추출(抽出)하고 분석(分析)을 한 결과(結果) 도입(導入)된 역전사 효소 유전자(遺傳子)의 전사(轉寫)가 고온(高溫)에서 증대(增大)되었다.

  • PDF

Control of Influenza: Live Vaccine Development

  • Seong, Baik-Lin
    • 대한약학회:학술대회논문집
    • /
    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
    • /
    • pp.149-150
    • /
    • 2002
  • Despite various efforts on improving vaccines and antivirals, influenza epidemics continue to afflict many people, causing widespread morbidity and mortality in the young and the elderly. Since the discovery of the unusual 'cap-stealing'mechanism of transcription, significant advances were made on molecular aspects of influenza gene regulation. This provides new insights for developing new antiviral compounds. Reverse genetic technologies have also been advanced for generating recombinant chimeric viruses suitable for designing live vaccine. (omitted)

  • PDF