• 제목/요약/키워드: Cell entry

검색결과 168건 처리시간 0.03초

An OTHBVS Cell Line Expresses the Human HBV Middle S Protein

  • Park, Sung-Gyoo;Guhung Jung
    • Journal of Microbiology
    • /
    • 제37권2호
    • /
    • pp.86-89
    • /
    • 1999
  • An OTHBVS cell line from HepG2 was established. This cell line stably expresses the human hepatitis B virus (HBV) middle S protein that includes the preS2 region which is important for HBV particle entry into the hepatocyte. To establish this cell line, the middle S open reading frame (ORF), with a promoter located in the 5' region and enhancer located in the 3' region, was cloned downstream from the metallothionine (MT) promoter of the OT1529 vector. In this vector, expression of the middle S protein was constructed to be regulated by its own promoter and enhancer. Expression of the large S protein which contains the preS1 region in addition to the middle S protein was designed to be regulated by the MT promoter. When extracts of OTHBVS cells were examined with an S protein detection kit (RPHA, Korea Green Cross Co.), an S protein was detected. Total mRNA of OTHBVS cell examined by northern blot analysis with an S ORF probe revealed small/middle S transcripts (2.1 kb). When the MT promoter was induced by Zn, large S transcripts (2.4 kb) were detected. The GP36 and GP33 middle S proteins were presumably detected, but large S proteins were not detected by immunostain analysis using anti-preS2 antibody.

  • PDF

Real-Time Temporal Dynamics of Bicistronic Expression Mediated by Internal Ribosome Entry Site and 2A Cleaving Sequence

  • Lee, Soomin;Kim, Jeong-Ah;Kim, Hee-Dae;Chung, Sooyoung;Kim, Kyungjin;Choe, Han Kyoung
    • Molecules and Cells
    • /
    • 제42권5호
    • /
    • pp.418-425
    • /
    • 2019
  • Multicistronic elements, such as the internal ribosome entry site (IRES) and 2A-like cleavage sequence, serve crucial roles in the eukaryotic ectopic expression of exogenous genes. For utilization of multicistronic elements, the cleavage efficiency and order of elements in multicistronic vectors have been investigated; however, the dynamics of multicistronic element-mediated expression remains unclear. Here, we investigated the dynamics of encephalomyocarditis virus (EMCV) IRES- and porcine teschovirus-1 2A (p2A)-mediated expression. By utilizing real-time fluorescent imaging at a minute-level resolution, we monitored the expression of fluorescent reporters bridged by either EMCV IRES or p2A in two independent cultured cell lines, HEK293 and Neuro2a. We observed significant correlations for the two fluorescent reporters in both multicistronic elements, with a higher correlation coefficient for p2A in HEK293 but similar coefficients for IRES-mediated expression and p2A-mediated expression in Neuro2a. We further analyzed the causal relationship of multicistronic elements by convergent cross mapping (CCM). CCM revealed that in all four conditions examined, the expression of the preceding gene causally affected the dynamics of the subsequent gene. As with the cross correlation, the predictive skill of p2A was higher than that of IRES in HEK293, while the predictive skills of the two multicistronic elements were indistinguishable in Neuro2a. To summarize, we report a significant temporal correlation in both EMCV IRES- and p2A-mediated expression based on the simple bicistronic vector and real-time fluorescent monitoring. The current system also provides a valuable platform to examine the dynamic aspects of expression mediated by diverse multicistronic elements under various physiological conditions.

Experimental Study on Impact Loads Acting on Free-falling Modified Wigley

  • Hong, Sa-Young;Kim, Young-Shik;Kyoung, Jo-Hyun;Hong, Seok-Won;Kim, Yong-Hwan
    • International Journal of Ocean System Engineering
    • /
    • 제2권3호
    • /
    • pp.151-159
    • /
    • 2012
  • The characteristics of an impact load and pressure were experimentally investigated. Drop tests were carried out using a modified Wigley with CB = 0.56. The vertical force, pressures, and vertical accelerations were measured. A 6-component load cell was used to measure the forces, piezo-electric sensors were used to capture the impact pressure, and strain-gauge type accelerometers were used to measure the vertical accelerations. A 50-kHz sampling rate was applied to capture the peak values. The repeatability of the measured data was confirmed and the basic characteristics of the impact load and pressure such as the linearity to the falling height were observed for all of the measurements. A simple formula was derived to extract the physical impact load from the measured force based on a simple mass-sensor-mass diagram, which was validated by comparing impact forces with existing data using the mathematical model of Faltinsen and Chezhian (2005). The effects of the elasticity of the model and change in acceleration during the water entry were investigated. It is interesting to observe that the impact loads occurred and reached peak values at the same time duration after water entry for all drop heights.

2사이클 기관 흡기 포오트의 가스 유동 특성에 관한 연구 (A study on the characteristics of gas flow in inlet port of 2 cycle engine)

  • 이창식
    • 대한기계학회논문집
    • /
    • 제11권5호
    • /
    • pp.725-730
    • /
    • 1987
  • 본 연구에서는 2사이클 기관의 흡입 포오트 부근의 가스 유동을 레이저 도플 러 유속계를 이용하여 측정하고 가스의 평균 속도, 속도 변동, 유속 벡터 등을 기관의 회전 속도, 실린더 헤드의 모양 및 실린더로의 흡입 속도 측정점의 위치 변화에 대하 여 비교 검토하고, 유동 특성을 고찰하였다.

ANKS1A-Deficiency Aberrantly Increases the Entry of the Protein Transport Machinery into the Ependymal Cilia

  • Haeryung Lee;Jiyeon Lee;Miram Shin;Soochul Park
    • Molecules and Cells
    • /
    • 제46권12호
    • /
    • pp.757-763
    • /
    • 2023
  • In this study, we examine whether a change in the protein levels for FOP in Ankyrin repeat and SAM domain-containing protein 1A (ANKS1A)-deficient ependymal cells affects the intraflagellar transport (IFT) protein transport system in the multicilia. Three distinct abnormalities are observed in the multicilia of ANKS1A-deficient ependymal cells. First, there were a greater number of IFT88-positive trains along the cilia from ANKS1A deficiency. The results are similar to each isolated cilium as well. Second, each isolated cilium contains a significant increase in the number of extracellular vesicles (ECVs) due to the lack of ANKS1A. Third, Van Gogh-like 2 (Vangl2), a ciliary membrane protein, is abundantly detected along the cilia and in the ECVs attached to them for ANKS1A-deficient cells. We also use primary ependymal culture systems to obtain the ECVs released from the multicilia. Consequently, we find that ECVs from ANKS1A-deficient cells contain more IFT machinery and Vangl2. These results indicate that ANKS1A deficiency increases the entry of the protein transport machinery into the multicilia and as a result of these abnormal protein transports, excessive ECVs form along the cilia. We conclude that ependymal cells make use of the ECV-based disposal system in order to eliminate excessively transported proteins from basal bodies.

플라스틱 세포치료제 보관용기의 극저온 환경 노출 시 구조적 변화에 대한 해석적 비교 (Analytical comparison of structural changes of plastic cell-based therapeutic drug storage containers when exposed to cryogenic environments)

  • 박정연;이동목;;이순례;윤길상
    • Design & Manufacturing
    • /
    • 제14권2호
    • /
    • pp.1-6
    • /
    • 2020
  • Recently, research and commercialization related to the field of cell-based therapeutic drug development has been actively conducted. In order to maintain cell viability and prevent contamination, refrigeration preservation devices, such as CRF (controlled rate freezer) or vapor type LN2 tanks have been developed. On the other hand, the storage container for liquid nitrogen tanks currently on sale minimizes the flow structure to prevent structural defects when stored in a liquid nitrogen tank having a high thermal conductivity than vapor nitrogen. If the cell-based treatment drug is stored in the gaseous LN2 tank as it is, the cell survival after thawing is greatly reduced. It was estimated that the existing storage container structure was a factor that prevented the rapid entry and circulation of gaseous nitrogen into the container. Therefore, this study intends to propose a new supercellular storage container model that can maintain the mechanical strength while maximizing the fluid flow structure. To this end, we estimated that the structural change of the storage container effects on the equivalent stress formed around the through-holes of them when exposed to a cryogenic environment using thermal-structural coupled field analysis. As a result of storage experiments in the gas phase tank of the cell-based therapeutic agent using the developed storage container, it was confirmed that the cell growth rate was improved from 66% to 77%, which satisfied the transportation standards of the FDA(Food and Drug Administration) cell-based therapeutic agent.

Two-Dimensional Ordination 분석법에 의한 제초제살초 Spectrum 분류와 효과적인 사용법 (A New Approach for Practical Classification of Herbicide and for Effective Use by Two-dimensional Ordination Analysis)

  • 김순철
    • 한국응용곤충학회지
    • /
    • 제22권2호
    • /
    • pp.147-159
    • /
    • 1983
  • In general, herbicides have been classified according to selectivity, mobility. time of application, methods of application, mode of action and chemical property and structure. However, there was no generally accepted classification system for practical use in the field. The primary processes affected by the majority of herbicides are the growth process through cell elongation and/or cell division, the photosynthetic process specifically the light reaction, the oxidative phosphorylation and the integrity of the membrane systems. The usual approach in the study of the mechanism by which herbicides kill or inhibit the growth of plants is to initially determine the morphological phototoxicity systems, The mechanism by which a herbicide kills a plant or suppresses its development is actually the resultant effect of primary and secondary(or side) effects. In most instances, the death of the plant is due to the secondary effects. To induce the desired response, a herbicide must be able to gain entry into the plants and once inside, to be transported within the plant to its site(s) of activity in concentrations great enough. Obstacles to the entry and movement of herbicides in plants are generally classified by leaf and soil obstacles, translocation obstacles and biochemical obstacles, and these obstacles are also strongly influenced by plant species and by environmental factors such as light, temperature, rainfall and relative humidity. And hence, in most instances, results obtained from laboratory or greenhous vary from those of field experiment. Author attempted to classify herbicides from the field experiment using the two-dimensional ordination analysis to obtain practical information for selecting effective herbicides or to choose effective herbicide combinations for increasing herbicidal efficacy or reducing the chemical cost. Based on this two-dimensional diagram, desired herbicides or combinations were selected and further investigated for the interaction effects whether these combinations are synergistic, additive or antagonistic. From the results, it was concluded that these new approach could possibly be give more comprehensive informations about effective use of herbicide than any other systems.

  • PDF

원격 조선소의 펨토셀 로컬 게이트웨이 구성과 패킷 오프로드 방식 제안 (A Proposal for a Femto-Cell Local Gateway and Packet Off-Loading for Remote Shipyards)

  • 이원희;민상원;정민아;이성로
    • 한국통신학회논문지
    • /
    • 제38C권4호
    • /
    • pp.387-393
    • /
    • 2013
  • 조선소는 다양한 용량 및 규격을 여러 사이트에서 제작하여 최종통합하기까지 여러 단계를 걸치게 되며 이들 사이의 정보공유가 필수적이며 최근 유무선통신망을 활용하여 단순 문자에서 동영상 정보를 주고받아 조선 산업의 스마트화가 이루어지고 있다. 본 논문에서는 원격에 위치한 조선소에 적용할 수 있는 LTE 펨토셀 로컬 게이트웨이 구성을 제안하고 상위 조선소와 중요정보를 주고받는 트래픽을 주고받기 위한 방안과 일반 트래픽을 분리하는 오프로드 방식을 제안하였다. 상위 조선소 메인 서버와 하위 원격 조선소 간에 정보와 일반 트래픽의 오프로드를 지원하기위해서 펨토셀 게이트웨이에서의 모드 전환, 오프로드 관련 정보 관리, 트리거로서의 메시지 정의, cache entry를 정의하였다 우리는 펨토셀 게이트웨이에서의 송수신 메시지 플로우와 state transition diagram을 이용하여 제안하는 방안의 기능 수행 여부를 확인하였다. 우리는 이동통신망과 광대역 인터넷 활용 기술을 조선 산업에 적용하여 조선소 산업 생산성 향상을 고려하였다.

Computer-aided drug design of Azadirachta indica compounds against nervous necrosis virus by targeting grouper heat shock cognate protein 70 (GHSC70): quantum mechanics calculations and molecular dynamic simulation approaches

  • Islam, Sk Injamamul;Saloa, Saloa;Mahfuj, Sarower;Islam, Md Jakiul;Jahan Mou, Moslema
    • Genomics & Informatics
    • /
    • 제20권3호
    • /
    • pp.33.1-33.17
    • /
    • 2022
  • Nervous necrosis virus (NNV) is a deadly infectious disease that affects several fish species. It has been found that the NNV utilizes grouper heat shock cognate protein 70 (GHSC70) to enter the host cell. Thus, blocking the virus entry by targeting the responsible protein can protect the fishes from disease. The main objective of the study was to evaluate the inhibitory potentiality of 70 compounds of Azadirachta indica (Neem plant) which has been reported to show potential antiviral activity against various pathogens, but activity against the NNV has not yet been reported. The binding affinity of 70 compounds was calculated against the GHSC70 with the docking and molecular dynamics (MD) simulation approaches. Both the docking and MD methods predict 4 (PubChem CID: 14492795, 10134, 5280863, and 11119228) inhibitory compounds that bind strongly with the GHSC70 protein with a binding affinity of -9.7, -9.5, -9.1, and -9.0 kcal/mol, respectively. Also, the ADMET (absorption, distribution, metabolism, excretion, and toxicity) properties of the compounds confirmed the drug-likeness properties. As a result of the investigation, it may be inferred that Neem plant compounds may act as significant inhibitors of viral entry into the host cell. More in-vitro testing is needed to establish their effectiveness.