• Title/Summary/Keyword: domain expression

Search Result 905, Processing Time 0.029 seconds

Soluble Expression and Purification of the Catalytic Domain of Human Vascular Endothelial Growth Factor Receptor 2 in Escherichia coli

  • Wei, Jia;Cao, Xiaodan;Zhou, Shengmin;Chen, Chao;Yu, Haijun;Zhou, Yao;Wang, Ping
    • Journal of Microbiology and Biotechnology
    • /
    • v.25 no.8
    • /
    • pp.1227-1233
    • /
    • 2015
  • Vascular endothelial growth factor (VEGF) plays a key role in angiogenesis through binding to its specific receptors, which mainly occurs to VEGF receptor 2 (VEGFR-2), a kinase insert domain-containing receptor. Therefore, the disruption of VEGFR-2 signaling provides a promising therapeutic approach for the treatment of cancer by inhibiting abnormal or tumorinduced angiogenesis. To explore this potential, we expressed the catalytic domain of VEGFR-2 (VEGFR-2-CD) as a soluble active kinase in Escherichia coli. The recombinant protein was purified and the VEGFR-2-CD activity was investigated. The obtained VEGFR-2-CD showed autophosphorylation activity and phosphate transfer activity comparable to the commercial enzyme. Furthermore, the IC50 value of known VEGFR-2 inhibitor was determined using the purified VEGFR-2-CD. These results indicated a possibility for functional and economical VEGFR-2-CD expression in E. coli to use for inhibitor screening.

Cognitive Domain of Problems in Korea Mathematics Textbooks (중학교 수학 교과서 문제의 인지적 영역 분석)

  • Ee, Ji Hye;Huh, Youjin;Shin, Minkyong;Huh, Nan
    • East Asian mathematical journal
    • /
    • v.35 no.4
    • /
    • pp.451-465
    • /
    • 2019
  • Textbooks are official materials so that these are the most frequently used teaching materials in school. The teacher constructs the lesson based on the contents of the textbook to achieve the learning goal. Thus, textbooks play an important role because the quality of the contents in textbooks affects the cognitive level of students. This study investigates the cognitive domain based on Bloom's Taxonomy of Educational Objectives(knowledge, understanding, application, analysis, synthesis and evaluation) of 'Values and Expression' in the mathematics textbook of the first grade of middle school reflecting the 2015 revised mathematics curriculum. We also looked at cognitive levels based on Bloom's Taxonomy of Educational Objectives. As a result, it was found that understanding was dominant in 'Values and Expression'. Also, although the problem of requiring a higher level of cognition is increasing as the unit finishes, there are still a high percentage of low level of problems.

Misexpression of AtTX12 encoding a Toll/interleukin-1 receptor domain induces growth defects and expression of defense-related genes partially independently of EDS1 in Arabidopsis

  • Song, Sang-Kee
    • BMB Reports
    • /
    • v.49 no.12
    • /
    • pp.693-698
    • /
    • 2016
  • In this study, a tissue-specific GAL4/UAS activation tagging system was used for the characterization of genes which could induce lethality when ubiquitously expressed. A dominant mutant exhibiting stunted growth was isolated and named defective root development 1-D (drd1-D). The T-DNA tag was located within the promoter region of AtTX12, which is predicted to encode a truncated nucleotide-binding leucine-rich repeat (NLR) protein, containing a Toll/interleukin-1 receptor (TIR) domain. The transcript levels of AtTX12 and defense-related genes were elevated in drd1-D, and the misexpression of AtTX12 recapitulated the drd1-D phenotypes. In the presence of ENHANCED DISEASE SUSCEPTIBILITY 1 (EDS1), a key transducer of signals triggered by TIR-type NLRs, a low-level of AtTX12 misexpression induced strong defective phenotypes including seedling lethality whereas, in the absence of EDS1, a high-level of AtTX12 misexpression induced weak growth defects like dwarfism, suggesting that AtTX12 might function mainly in an EDS1-dependent and partially in an EDS1-independent manner.

Dpp Represses eagle Expression at Short-Range, but Can Repress Its Expression at a Long-Range via EGFR Signal Repression

  • Kim, Se Young;Jung, Keuk Il;Kim, Sang Hee;Jeon, Sang-Hak
    • Molecules and Cells
    • /
    • v.26 no.6
    • /
    • pp.576-582
    • /
    • 2008
  • Nervous system development takes place after positional information has been established along the dorsal-ventral (D/V) axis. The initial subdivision provided by a gradient of nuclear dorsal protein is maintained by the zygotic genes expressed along the D/V axis. In this study, an investigation was conducted to determine the range of Dpp function in repressing the expression of eagle (eg) that is present in intermediate neuroblasts defective (ind) and muscle specific homeobox (msh) gene domain. eg is expressed in neuroblast (NB) 2-4, 3-3 and 6-4 of the msh domain, and NB7-3 of the ind domain at the embryonic stage 11. In decapentaplegic (dpp) loss-of-function mutant embryos, eg was ectopically expressed in the dorsal region, while in dpp gain-of-function mutants produced by sog or sca-GAL4/UAS-dpp, eg was repressed by Dpp. It is worthy of note that Dpp produced from sim;;dpp embryos showed that Dpp could function at long range. However, Dpp produced from en-GAL4/UAS-dpp or wg-GAL4/UAS-dpp primarily acted at short-range. This result demonstrated that this discrepancy seems to be due to the repression of Dpp to EGFR signaling in sim;;dpp embryos. Taken together, these results suggest that Dpp signaling works at short-range, but can function indirectly at long-range by way of repression of EGFR signaling during embryonic neurogenesis.

Gene Transfer and Gene Expression of Novel Recombinant Baculovirus Vector System (새로운 재조합 베큘로바이러스벡터의 유전자전이와 유전자발현)

  • Sa, Young-Hee;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
    • /
    • 2013.10a
    • /
    • pp.946-948
    • /
    • 2013
  • Several baculovirus vector systems recombined with coding genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD) were constructed. These recombinant baculovirus vector systems were applied into human foreskin fibroblast cells and compared the effects of gene transfer and gene expression of these recombinant baculovirus vector systems with control vector system. From this study, it showed that these novel recombinant baculovirus vector systems were superior efficacy to control vector system in view of gene transfer and gene expression.

  • PDF

Expression of Gal4-VP16 and Gal4-DNA binding domain under the control of the T lymphocyte-specific lck proximal promoter in transgenic mice

  • Ryu, Chun-Jeih;Whitehurst, Charles E.;Chen, Jianzhu
    • BMB Reports
    • /
    • v.41 no.8
    • /
    • pp.575-580
    • /
    • 2008
  • Thymocyte-specific transcriptional regulatory systems can be used to better understand the relationship between transcription and V(D)J recombination during early T cell development. In this study, we generated transgenic mice expressing the transactivator Gal4-VP16 or the Gal4 DNA binding domain (Gal4-DBD) under the control of the lck proximal promoter, which is only active in immature thymocytes. From these studies Gal4-VP16 and Gal4-DBD expression was shown to significantly alter thymic cellularity and differentiation without significantly changing the $CD3^+$ thymocyte distribution. Furthermore, the presence of Gal4-VP16 or Gal4-DBD in the transgenic thymocytes retarded the mobility of the Gal4 DNA binding motif as determined by a gel mobility shift assay, suggesting that the developmental alteration did not affect the functional property of the transgenic proteins. These results indicated that lck promoter-driven Gal4-VP16 or Gal4-DBD expression did not affect $CD3^+$ mature thymocytes, thus this system can be applied to study transcriptional regulation of transresponder genes in bigenic mouse model thymocytes.

Gene Transfer and Expression of Newly Reconstructed Baculovirus Vectors (재조성된 베큘로바이러스 벡터의 유전자 전이와 발현)

  • Kim, Ji-Young;Kim, Hyun Joo;Sa, Young-Hee;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
    • /
    • 2016.10a
    • /
    • pp.923-926
    • /
    • 2016
  • Baculovirus vectors were reconstructed using cytomegalovirus (CMV) promoter, polyhedron promoter, vesicular stomatitis virus G (VSVG), enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD) genes. These reconstructed vector was transfected into various cell lines and tissues. We compared this reconstructed vector with other control vectors in view of gene transfer and gene expression. In conclusion, we confirmed that gene transfer and expression of these reconstructed vectors was higher efficient than any other control vector.

  • PDF

Efficacy of Gene Transfer and Expression of Recombinanat Baculovirus Vector System (재조합 베큘로바이러스벡터의 유전자전달과 발현의 효과)

  • Sa, Young-Hee;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
    • /
    • 2014.05a
    • /
    • pp.813-815
    • /
    • 2014
  • Novel baculovirus vector systems including genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD) were constructed. These recombinant baculovirus vector systems were transfected into diverse cells of 293T, HepG2, HFF, and Hur7 cells and compared the effects of gene transfer and expression of these vector systems with control vector. From the result, we confirmed that these recombinant baculovirus vector systems were more excellent than control vector in efficacy of gene transfer and expression.

  • PDF

Cytosolic domain regulates the calcium sensitivity and surface expression of BEST1 channels in the HEK293 cells

  • Kwon Woo Kim;Junmo Hwang;Dong-Hyun Kim;Hyungju Park;Hyun-Ho Lim
    • BMB Reports
    • /
    • v.56 no.3
    • /
    • pp.172-177
    • /
    • 2023
  • BEST family is a class of Ca2+-activated Cl- channels evolutionary well conserved from bacteria to human. The human BEST paralogs (BEST1-BEST4) share significant amino acid sequence homology in the N-terminal region, which forms the transmembrane helicases and contains the direct calcium-binding site, Ca2+-clasp. But the cytosolic C-terminal region is less conserved in the paralogs. Interestingly, this domain-specific sequence conservation is also found in the BEST1 orthologs. However, the functional role of the C-terminal region in the BEST channels is still poorly understood. Thus, we aimed to understand the functional role of the C-terminal region in the human and mouse BEST1 channels by using electrophysiological recordings. We found that the calcium-dependent activation of BEST1 channels can be modulated by the C-terminal region. The C-terminal deletion hBEST1 reduced the Ca2+-dependent current activation and the hBEST1-mBEST1 chimera showed a significantly reduced calcium sensitivity to hBEST1 in the HEK293 cells. And the C-terminal domain could regulate cellular expression and plasma membrane targeting of BEST1 channels. Our results can provide a basis for understanding the C-terminal roles in the structure-function of BEST family proteins.

Expression and characterization of transmembrane and coiled-coil domain family 3

  • Sohn, Wern-Joo;Kim, Jae-Young;Kim, Dongbum;Park, Jeong-A;Lee, Younghee;Kwon, Hyung-Joo
    • BMB Reports
    • /
    • v.49 no.11
    • /
    • pp.629-634
    • /
    • 2016
  • Transmembrane and coiled-coil domain family 3 (TMCC3) has been reported to be expressed in the human brain; however, its function is still unknown. Here, we found that expression of TMCC3 is higher in human whole brain, testis and spinal cord compared to other human tissues. TMCC3 was expressed in mouse developing hind brain, lung, kidney and somites, with strongest expression in the mesenchyme of developing tongue. By expression of recombinant TMCC3 and its deletion mutants, we found that TMCC3 proteins self-assemble to oligomerize. Immunostaining and confocal microscopy data revealed that TMCC3 proteins are localized in endoplasmic reticulum through transmembrane domains. Based on immunoprecipitation and mass spectroscopy data, TMCC3 proteins associate with TMCC3 and 14-3-3 proteins. This supports the idea that TMCC3 proteins form oligomers and that 14-3-3 may be involved in the function of TMCC3. Taken together, these results may be useful for better understanding of uncharacterized function of TMCC3.