• 제목/요약/키워드: Functional characterization

검색결과 793건 처리시간 0.03초

Identification and Functional Characterization of the GALACTINOL SYNTHASE (MoGolS1) Gene in Melissa officinalis Plants

  • Kim, Jun-Hyeok;Hossain, Acktar Mohammad;Kim, Na-Hyun;Lee, Dong-Ho;Lee, Ho-Joung
    • Journal of Applied Biological Chemistry
    • /
    • 제54권4호
    • /
    • pp.244-251
    • /
    • 2011
  • Galactinol and rafinose accumulation in plants is associated with stressful environmental conditions such as cold, heat, or dehydration by the action of galactinols synthase (GolS) in the raffinose family of oligosaccharides biosynthetic pathway from UDP-galactose. Moreover, several reports mentioned that GolS transcription is up regulated by various environmental stresses like cold, heat, dehydration. Therefore, to determine whether MoGolS1 was induced with the abiotic stress we analyzed the expression pattern of the gene under various abiotic stresses like heat, cold, abscisic acid, sucrose and salt concentration in the lemon balm plants grown in standard MS medium. The MoGolS1 gene was 981-bp in length encoding 326 amino acids in its sequence and shared 77 and 76% sequence similarity with Arabidopsis thaliana galactinol synthase4 (AtGolS4) and AtGolS1 genes respectively. The MoGolS1 gene was strongly expressed by the abiotic stress induced by sucrose, ABA or heat shock. It was also expressed in responses to cold, Identification and Functional Characterization of the GALACTINOL SYNTHASgene induction with various stresses may be possible for itscrucial function in abiotic stress tolerance in plants, providing a good engineering target for genetic engineering.

배추 유래 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase 관련 Brmecp 유전자의 발현 및 분자적 특성 (Molecular and functional characterization of a Brmecp gene encoding 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase from Brassica rapa)

  • 정유진;최장선;선주남;노일섭;조용구;강권규
    • Journal of Plant Biotechnology
    • /
    • 제39권3호
    • /
    • pp.189-195
    • /
    • 2012
  • In plants, the fifth step of the plastidial 2-Cmethyl-D-erythritol 4-phosphate (MEP) pathway is catalyzed by 2-C-Methyl-D-erythritol 2,4-cyclodiphosphate synthase (MECP; EC: 4. 6. 1. 12), an enzyme proposed to play a key role in the regulation of isoprenoid biosynthesis. Here we report the isolation and functional characterization of a 823 bp Brassica rapa MECP (Brmecp) cDNA encoding a deduced polypeptide of 230 amino acid residues. Transcription levels of Brmecp were two-fold higher in petal compared to leaves. In addition, Brmecp expression in cabbage seedlings treated with ABA, $H_2O_2$ and drought was higher than control seedlings. These results were consistent with changes in chlorophyll contents in transgenic Arabidopsis. Thus, the Brmecp may contribute to the production of primary (chlorophylls and carotenoids) isoprenoid end-products in chloroplasts.

Rubus속 식물에 존재하는 $19{\alpha}-Hydroxyursane-type$ Triterpenoid의 특성과 이용 (Chemical Characterization and Utilization of $19{\alpha}-Hydroxyursane-type$ Triterpenoids in Rubus species)

  • 정현주;남정환;임상철;김원배;박희준
    • 한국자원식물학회지
    • /
    • 제19권5호
    • /
    • pp.563-572
    • /
    • 2006
  • The plant Rubus species (Rosaceae) mainly contains $19{\alpha}-hydroxyurane-type$ triterpenoids $(19{\alpha}-HUT)$ as bioactive components. Available functional food includes blackberry (the fruit of thornless Rubus sp.), red raspberry (R. idaeus) and black raspberry (R. occidentalis). However, the fruit of R. coreanus, which is used in Korea as a functional food, substitutes black raspberry. Rubi Fructus, which has been traditionally used as an oriental medicinal drug, designates only unripe fruit of R. coreanus but not its ripe fruit which indicates that it needs high content of $19{\alpha}-HUT$ as a crude drug. Throughout our experiment, we found that ripe fruits contain very little amount of $19{\alpha}-HUT$ when compared to unripe fruits. In addition, various and rich $19{\alpha}-HUT$ has been reported from Rubus species. The most common structure of $19{\alpha}-HUT$ of Rubus species, euscaphic acid or tormentic acid with $3{\alpha}-OH$ or $3{\beta}-OH$, respectively, can be glycosylated or oxidized to produce a number of $19{\alpha}-HUTs $as euscaphic acid and tormentic acid derivatives and even esterified to form dimeric triterpenoids. In this review, the bioactivity and biosynthetic pathway and chemical characterization of $19{\alpha}-HUTs$ found in Rubus species are discussed.

균류 Coprinus cinereus에서 DNA 회복에 관여하는 RAD4 유사유전자의 분리와 특성 (Characterization of RAD4 Homologous Gene from Coprinus cinereus)

  • Choi, In-Soon
    • 생명과학회지
    • /
    • 제13권4호
    • /
    • pp.522-528
    • /
    • 2003
  • 본 연구는 출아형 효모 Saccharomyces cerevisiae에서 자외선의 상해 시 이를 정상으로 회복시키는 절제회복 (excision repair) 유전자로 알려진 RADA4의 특성 규명을 위하여 균류 Coprinus cinereus에서 이와 유사한 유전자를 분리하였다. RAD4 유사 유전자를 분리하기 위하여 균류 C. cinereus의 염색체 DNA를 전기영동하여 분리한 다음 효모 RAD4 DNA를 probe로하여 이와 hybridization하였다. 이 결과 RAD4 유사 유전자는 3.2 kb의 insert DNA를 갖고 있었다. 또한 Southern hybridization으로 이 유사 유전자는 fungus C. cinereus의 염색체에 존재함을 확인하였다. 분리한 RAD4 유사 유전자의 전사체 크기는 2.5 kb 였으며, 자외선의 상해 시 전혀 'inducibility가 없음을 Northern hybridization으로 확인하였다. 또한 유사유전자 부분을 삭제하였을 때 이 부분이 없는 세포는 전혀 생존을 못하였다. 이 결과 분리한 RAD4 유사유전자는 세포의 생존에 관여함을 알 수 있었다.

Functional Characterization of Pharmcogenetic Variants of Human Cytochrome P450 2C9 in Korean Populations

  • Cho, Myung-A;Yoon, Jihoon G.;Kim, Vitchan;Kim, Harim;Lee, Rowoon;Lee, Min Goo;Kim, Donghak
    • Biomolecules & Therapeutics
    • /
    • 제27권6호
    • /
    • pp.577-583
    • /
    • 2019
  • Human cytochrome P450 2C9 is a highly polymorphic enzyme that is required for drug and xenobiotic metabolism. Here, we studied eleven P450 2C9 genetic variants-including three novel variants F69S, L310V, and Q324X-that were clinically identified in Korean patients. P450 2C9 variant enzymes were expressed in Escherichia coli and their bicistronic membrane fractions were prepared The CO-binding spectra were obtained for nine enzyme variants, indicating P450 holoenzymes, but not for the M02 (L90P) variant. The M11 (Q324X) variant could not be expressed due to an early nonsense mutation. LC-MS/MS analysis was performed to measure the catalytic activities of the P450 2C9 variants, using diclofenac as a substrate. Steady-state kinetic analysis revealed that the catalytic efficiency of all nine P450 2C9 variants was lower than that of the wild type P450 2C9 enzyme. The M05 (R150L) and M06 (P279T) variants showed high $k_{cat}$ values; however, their $K_m$ values were also high. As the M01 (F69S), M03 (R124Q), M04 (R125H), M08 (I359L), M09 (I359T), and M10 (A477T) variants exhibited higher $K_m$ and lower $k_{cat}$ values than that of the wild type enzyme, their catalytic efficiency decreased by approximately 50-fold compared to the wild type enzyme. Furthermore, the novel variant M07 (L310V) showed lower $k_{cat}$ and $K_m$ values than the wild type enzyme, which resulted in its decreased (80%) catalytic efficiency. The X-ray crystal structure of P450 2C9 revealed the presence of mutations in the residues surrounding the substrate-binding cavity. Functional characterization of these genetic variants can help understand the pharmacogenetic outcomes.

A genome-wide approach to the systematic and comprehensive analysis of LIM gene family in sorghum (Sorghum bicolor L.)

  • Md. Abdur Rauf Sarkar;Salim Sarkar;Md Shohel Ul Islam;Fatema Tuz Zohra;Shaikh Mizanur Rahman
    • Genomics & Informatics
    • /
    • 제21권3호
    • /
    • pp.36.1-36.19
    • /
    • 2023
  • The LIM domain-containing proteins are dominantly found in plants and play a significant role in various biological processes such as gene transcription as well as actin cytoskeletal organization. Nevertheless, genome-wide identification as well as functional analysis of the LIM gene family have not yet been reported in the economically important plant sorghum (Sorghum bicolor L.). Therefore, we conducted an in silico identification and characterization of LIM genes in S. bicolor genome using integrated bioinformatics approaches. Based on phylogenetic tree analysis and conserved domain, we identified five LIM genes in S. bicolor (SbLIM) genome corresponding to Arabidopsis LIM (AtLIM) genes. The conserved domain, motif as well as gene structure analyses of the SbLIM gene family showed the similarity within the SbLIM and AtLIM members. The gene ontology (GO) enrichment study revealed that the candidate LIM genes are directly involved in cytoskeletal organization and various other important biological as well as molecular pathways. Some important families of regulating transcription factors such as ERF, MYB, WRKY, NAC, bZIP, C2H2, Dof, and G2-like were detected by analyzing their interaction network with identified SbLIM genes. The cis-acting regulatory elements related to predicted SbLIM genes were identified as responsive to light, hormones, stress, and other functions. The present study will provide valuable useful information about LIM genes in sorghum which would pave the way for the future study of functional pathways of candidate SbLIM genes as well as their regulatory factors in wet-lab experiments.

테오필린 분자 날인 고분자의 합성 및 특성 (Synthesis and Characterization of Theophylline Molecularly Imprinted Polymers)

  • 유호식;김범수;김대수
    • 폴리머
    • /
    • 제32권2호
    • /
    • pp.138-142
    • /
    • 2008
  • 분자 날인 기술은 표적 분자에 대해 높은 선택도를 갖는 합성 재료를 제조하기 위한 효과적인 방법이다. 본 연구에서는 주형 분자로 테오필린(theophylline)을, 가교제로 폴리에스터-아크릴레이트 수지를 사용하여 UV 중합을 통해 분자 날인 고분자(MIP)를 합성하였다. 기능성 단량체 종류가 MIP의 성능에 미치는 영향을 알아보기 위해, 메타크릴산(mathacrylic acid), 아크릴산(acrylic acid), 그리고 아크릴 아미드(acrylic amide)를 기능성 단량체로 각각 사용하여 MIP를 합성하였다. MIP는 비날인 고분자(NIP)보다 테오필린에 대해 훨씬 더 높은 재결합 능력을 보였다. 메타크릴산을 사용하여 합성한 MIP는 가장 높은 재결합 능력을 보였다. MIP의 선택도는 테오필린과 분자구조가 유사한 카페인(caffeine) 용액을 사용하여 조사하였다. 클로로포름보다 극성인 증류수를 용매로 사용하였을 경우 MIP의 테오필린 재결합 성능은 감소하였다.

Synthesis and Characterization of Red Electrophosphorescent Polymers Containing Pendant Iridium(III) Complex Moieties

  • Xu, Fei;Mi, Dongbo;Bae, Hong Ryeol;Suh, Min Chul;Yoon, Ung Chan;Hwang, Do-Hoon
    • Bulletin of the Korean Chemical Society
    • /
    • 제34권9호
    • /
    • pp.2609-2615
    • /
    • 2013
  • A series of fluorene-carbazole copolymers containing the pendant phosphor chromophore $Ir(absn)_2(acac)$ (absn: 2-(1-naphthyl)benzothiazole; acac: acetylacetone) were designed and synthesized via Yamamoto coupling. In the film state, these copolymers exhibited absorption and emission peaks at approximately 389 and 426 nm, respectively, which originated from the fluorene backbone. However, in electroluminescent (EL) devices, a significantly red-shifted emission at approximately 611 nm was observed, which was attributed to the pendant iridium(III) complex. Using these copolymers as a single emission layer, polymer light-emitting devices with ITO/PEDOT:PSS/polymer:DNTPD/TmPyPb/LiF/Al configurations exhibited a saturated red emission at 611 nm. The attached iridium(III) complex had a significant effect on the EL performance. A maximum luminous efficiency of 0.85 cd/A, maximum external quantum efficiency of 0.77, maximum power efficiency of 0.48 lm/W, and maximum luminance of 883 $cd/m^2$ were achieved from a device fabricated with the copolymer containing the iridium(III) complex in a 2% molar ratio.

Synthesis and Characterization of Biodegradable Thermo- and pH-Sensitive Hydrogels Based on Pluronic F127/Poly($\varepsilon$-caprolactone) Macromer and Acrylic Acid

  • Zhao, Sanping;Cao, Mengjie;Wu, Jun;Xu, Weilin
    • Macromolecular Research
    • /
    • 제17권12호
    • /
    • pp.1025-1031
    • /
    • 2009
  • Several kinds of biodegradable hydrogels were prepared via in situ photopolymerization of Pluronic F127/poly($\varepsilon$-caprolactone) macromer and acrylic acid (AA) comonomer in aqueous medium. The swelling kinetics measurements showed that the resultant hydrogels exhibited both thermo- and pH-sensitive behaviors, and that this stimuli-responsiveness underwent a fast reversible process. With increasing pH of the local buffer solutions, the pH sensitivity of the hydrogels was increased, while the temperature sensitivity was decreased. In vitro hydrolytic degradation in the buffer solution (pH 7.4, $37^{\circ}C$), the degradation rate of the hydrogels was greatly improved due to the introduction of the AA comonomer. The in vitro release profiles of bovine serum albumin (BSA) in-situ embedded into the hydrogels were also investigated: the release mechanism of BSA based on the Peppas equation was followed Case II diffusion. Such biodegradable dual-sensitive hydrogel materials may have more advantages as a potentially interesting platform for smart drug delivery carriers and tissue engineering scaffolds.

Functional Abnormalities of HERG Mutations in Long QT Syndrome 2 (LQT2)

  • Hiraoka, Masayasu
    • The Korean Journal of Physiology and Pharmacology
    • /
    • 제5권5호
    • /
    • pp.367-371
    • /
    • 2001
  • The chromosome 7-linked long QT syndrome (LQT2) is caused by mutations in the human ether-a- go-go-related gene (HERG) that encodes the rapidly activating delayed rectifier $K^+$ current, $I_{Kr},$ in cardiac myocytes. Different types of mutations have been identified in various locations of HERG channel. One of the mechanisms for the loss of normal channel function is due to membrane trafficking of channel protein. The decreased channel function in some deletion mutants appears to be due to loss of coupling with wild type HERG to form the functional channel as the tetramer. Most of missense mutants with few exceptions could interact with wild type HERG to form functional tetramer and caused dominant negative suppression with co-injection with wild type HERG showing variable effects on current amplitude, voltage dependence, and kinetics of activation and inactivation. Two missense mutants at pore regions of HERG found in Japanese LQT2 (A614V and V630L) showed accentuated inward rectification due to a negative shift in steady-state inactivation and fast inactivation. One mutation in S4 region (R534C) produced a negative shift in current activation, indicating the S4 serving as the voltage sensor and accelerated deactivation. The C-terminus mutation, S818L, could not express the current by mutant alone and did not show dominant negative suppression with co-injection of equal amount of wild type cRNA. Co-injection of excess amount of mutant with wild type produced dominant negative suppression with a shift in voltage dependent activation. Therefore, multiple mechanisms are involved in different mutations and functional abnormality in LQT2. Further characterization with the interactions between various mutants in HERG and the regulatory subunits of the channels (MiRP1 and minK) is to be clarified.

  • PDF