• Title/Summary/Keyword: mutant mice

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Construction and immunization with double mutant ΔapxIBD Δpnp forms of Actinobacillus pleuropneumoniae serotypes 1 and 5

  • Dao, Hoai Thu;Truong, Quang Lam;Do, Van Tan;Hahn, Tae-Wook
    • Journal of Veterinary Science
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    • 제21권2호
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    • pp.20.1-20.13
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    • 2020
  • Actinobacillus pleuropneumoniae (APP) causes a form of porcine pleuropneumonia that leads to significant economic losses in the swine industry worldwide. The apxIBD gene is responsible for the secretion of the ApxI and ApxII toxins and the pnp gene is responsible for the adaptation of bacteria to cold temperature and a virulence factor. The apxIBD and pnp genes were deleted successfully from APP serotype 1 and 5 by transconjugation and sucrose counter-selection. The APP1ΔapxIBDΔpnp and APP5ΔapxIBDΔpnp mutants lost hemolytic activity and could not secrete ApxI and ApxII toxins outside the bacteria because both mutants lost the ApxI- and ApxII-secreting proteins by deletion of the apxIBD gene. Besides, the growth of these mutants was defective at low temperatures resulting from the deletion of pnp. The APP1ΔapxIBDΔpnp and APP5ΔapxIBDΔpnp mutants were significantly attenuated compared with wild-type ones. However, mice vaccinated intraperitoneally with APP5ΔapxIBDΔpnp did not provide any protection when challenged with a 10-times 50% lethal dose of virulent homologous (APP5) and heterologous (APP1) bacterial strains, while mice vaccinated with APP1ΔapxIBDΔpnp offered 75% protection against a homologous challenge. The ΔapxIBDΔpnp mutants were significantly attenuated and gave different protection rate against homologous virulent wild-type APP challenging.

Acetobacter pasteurianus IFO 13751-5 변이주가 생산하는 다당류의 항암효과 (Antitumor Effect of Polysaccharide Produced from a Mutant of Acetobacter pasteurianus IFO 13751-5)

  • 김동석;류병호
    • 한국식품과학회지
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    • 제23권4호
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    • pp.405-409
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    • 1991
  • 다당류의 화학적인 조성은 다당류의 함량이 85.82% 이며, 이를 구성하는 단당류는 glucose의 함량이 79.60%로 가장 많고, 단백질의 함량은 2.07%이며, 구성 아미노산의 조성은 cystein, aspartic acid, valine, glutamic acid가 주요 아미노산이었다. 초산균에서 생산된 다당류가 Sarcoma 180에 대한 고형암 성장 저지율은 50 mg/kg의 투여시 64.96%의 가장 우수한 저지효과를 나타내었다. 수명 연장 효과는 50 mg/kg의 투여시 28.91%이었으며, 복수형암에 대해서는 별 높은 효과를 보이지 않았다. In vivo에서의 항암효과를 근거로 하여 in vitro에서는 Sarcoma 180에 대한 직접적 세포독성 작용은 거의 없었다.

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Connecting the dots between SHP2 and glutamate receptors

  • Ryu, Hyun-Hee;Kim, Sun Yong;Lee, Yong-Seok
    • The Korean Journal of Physiology and Pharmacology
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    • 제24권2호
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    • pp.129-135
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    • 2020
  • SHP2 is an unusual protein phosphatase that functions as an activator for several signaling pathways, including the RAS pathway, while most other phosphatases suppress their downstream signaling cascades. The physiological and pathophysiological roles of SHP2 have been extensively studied in the field of cancer research. Mutations in the PTPN11 gene which encodes SHP2 are also highly associated with developmental disorders, such as Noonan syndrome (NS), and cognitive deficits including learning disabilities are common among NS patients. However, the molecular and cellular mechanism by which SHP2 is involved in cognitive functions is not well understood. Recent studies using SHP2 mutant mice or pharmacological inhibitors have shown that SHP2 plays critical role in learning and memory and synaptic plasticity. Here, we review the recent studies demonstrating that SHP2 is involved in synaptic plasticity, and learning and memory, by the regulation of the expression and/or function of glutamate receptors. We suggest that each cell type may have distinct paths connecting the dots between SHP2 and glutamate receptors, and these paths may also change with aging.

Characterization of the cellular localization of C4orf34 as a novel endoplasmic reticulum resident protein

  • Jun, Mi-Hee;Jun, Young-Wu;Kim, Kun-Hyung;Lee, Jin-A;Jang, Deok-Jin
    • BMB Reports
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    • 제47권10호
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    • pp.563-568
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    • 2014
  • Human genome projects have enabled whole genome mapping and improved our understanding of the genes in humans. However, many unknown genes remain to be functionally characterized. In this study, we characterized human chromosome 4 open reading frame 34 gene (hC4orf34). hC4orf34 was highly conserved from invertebrate to mammalian cells and ubiquitously expressed in the organs of mice, including the heart and brain. Interestingly, hC4orf34 is a novel ER-resident, type I transmembrane protein. Mutant analysis showed that the transmembrane domain (TMD) of hC4orf34 was involved in ER retention. Overall, our results indicate that hC4orf34 is an ER-resident type I transmembrane protein, and might play a role in ER functions including $Ca^{2+}$ homeostasis and ER stress.

Association of MC4R Gene Polymorphisms with Growth and Body Composition Traits in Chicken

  • Li, Chun-Yu;Li, Hui
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권6호
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    • pp.763-768
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    • 2006
  • Genetic and pharmacological studies in mice have demonstrated a complementary role for the melanocortin 4 receptor (MC4R) in the control of food intake, energy balance and body weight. This study was designed to investigate the associations of a MC4R gene polymorphism on chicken growth and body composition traits in broiler lines divergently selected for abdominal fat. A SNP (G54C) was found in CDS region of chicken MC4R gene. The analysis of the least squares and variance revealed a significant association between the G54C SNP and BW, CW and SL at 7 wk of age, and there were significant differences in different genotypes (p<0.05). The results from protein secondary structure prediction and tertiary structure prediction showed that it appeared a helix in $13^{th}$ amino acid and two strands at $14^{th}$ and $15^{th}$ amino acid in mutant protein, respectively. It maybe induce the change of the activity or function of MC4R gene in poultry.

Characterization and Expression Pattern of Myostatin in the Rockfish, Sebastes schlegeli

  • Lee, Sang-Beum;Kim, Yong-Soo;Jin, Hyung-Joo
    • Fisheries and Aquatic Sciences
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    • 제10권2호
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    • pp.60-67
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    • 2007
  • Myostatin (MSTN; also known as GDF8) is a member of the transforming growth factor ${\beta}-superfamily$ of proteins. MSTN negatively regulates mammalian skeletal muscle growth and development by inhibiting myoblast proliferation. Mice and cattle possessing mutant MSTN alleles display a 'double muscling' phenotype characterized by extreme skeletal muscle hypertrophy and/or hyperplasia. We isolated the full-length cDNA of a novel MSTN gene from S. schlegeli muscle tissue and examined its expression pattern in various tissues. The full-length gene (GenBank DQ423474) consists of 1941bp with an open reading frame of 1134 bp, encoding 377 amino acids that show 62-92% amino acid similarity to other vertebrate MSTNs. The predicted protein contains a conserved proteolytic cleavage site (RXRR) and nine conserved cysteine residues at the C terminus. RT-PCR revealed that the unprocessed and prodomain myostatin mRNAs were predominantly present in muscle, with limited expression in other tissues. However, the mature myostatin mRNA was highly expressed in brain and muscle, intermediately expressed in the gills, intestine, heart, and kidney, and weakly expressed in the liver and spleen.

An embryoprotective role for glucose-6-phosphate dehydrogenase in developmental oxidative stress and chemical teratogenesis

  • Nicol, Christopher J.;Zielenski, Julian;Tsui, Lap-Chee;Wells, Peter G.
    • 한국환경성돌연변이발암원학회:학술대회논문집
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    • 한국환경성돌연변이발암원학회 2002년도 Current Trends in Toxicological Sciences
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    • pp.48-64
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    • 2002
  • The primary recognized health risk from common deficiencies in glucose-6-phosphate dehydrogenase (G6PD), a cytoprotective enzyme for oxidative stress, is red blood cell hemolysis. Here we show that litters from untreated pregnant mutant mice with a hereditary G6PD deficiency had increased prenatal (fetal resorptions) and postnatal death. When treated with the anticonvulsant drug phenytoin, a human teratogen that is commonly used in pregnant women and causes embryonic oxidative stress, G6PD-deficient dams had higher embryonic DNA oxidation and more fetal death and birth defects. The reported G6PD gene mutation was confirmed and used to genotype fetal resorptions, which were primarily G6PD deficient. This is the first evidence that G6PD is a developmentally critical cytoprotective enzyme for both endogenous and xenobiotic-initiated embryopathic oxidative stress and DNA damage. G6PD deficiencies accordingly may have a broader biological relevance as important determinants of infertility, in utero and postnatal death, and teratogenesis.-Nicol, C. J., Zielenski, J., Tsui, L.-C., Wells, P. G. An embryoprotective role for glucose-6-phosphate dehydrogenase in developmental oxidative stress and chemical teratogenesis.

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Pneumococcal Δpep27 Immunization Attenuates TLRs and NLRP3 Expression and Relieves Murine Ovalbumin-Induced Allergic Rhinitis

  • Yu, Jae Ik;Kim, Ji-Hoon;Nam, Ki-El;Lee, Wonsik;Rhee, Dong-Kwon
    • Journal of Microbiology and Biotechnology
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    • 제32권6호
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    • pp.709-717
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    • 2022
  • Allergic rhinitis (AR), one of the most common inflammatory diseases, is caused by immunoglobulin E (IgE)-mediated reactions against inhaled allergens. AR involves mucosal inflammation driven by type 2 helper T (Th2) cells. Previously, it was shown that the Streptococcus pneumoniae pep27 mutant (Δpep27) could prevent and treat allergic asthma by reducing Th2 responses. However, the underlying mechanism of Δpep27 immunization in AR remains undetermined. Here, we investigated the role of Δpep27 immunization in the development and progression of AR and elucidated potential mechanisms. In an ovalbumin (OVA)-induced AR mice model, Δpep27 alleviated allergic symptoms (frequency of sneezing and rubbing) and reduced TLR2 and TLR4 expression, Th2 cytokines, and eosinophil infiltration in the nasal mucosa. Mechanistically, Δpep27 reduced the activation of the NLRP3 inflammasome in the nasal mucosa by down-regulating the Toll-like receptor signaling pathway. In conclusion, Δpep27 seems to alleviate TLR signaling and NLRP3 inflammasome activation to subsequently prevent AR.

Helicobacter pylori에서 metronidazole 내성이 미치는 생물학적 영향 (Biological Effect of Metronidazole Resistance in Helicobacter Pylori)

  • 이진혁;이선미;남원희;김은실;김진호;강호영;정진용
    • 생명과학회지
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    • 제15권6호
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    • pp.955-960
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    • 2005
  • 메트로니다졸은 인체 위장질환의 원인균인 헬리코박터 파일로리를 박별하기위해 처방하는 주요 약제이다 그러나 인체로부터 분리한 헬리코박터 파일로리 균주는 메트로니다졸에 내성을 가지는 경우가 일반적이며, 이러한 내성원인은 이 균주의 염색체 상에 존재하는 두 종류의 nitroreductase 유전자인 rdxA와frxA 유전자가 비활성화됨에 따라 유발된다. 본 연구에서는 헬리코박터 파일로리 균에서 rdxA 유전자에 변이를 도입하여 메트로니다졸에 내성을 가지는 균주를 구축하여, 메트로니다졸 내성이 균주에 미치는 생물학적 영향을 관찰하고자 하였다. In vitro상에서 메트로니다졸 내성균주는 대조균과 비교하여 exponential phase에서는 거의 차이 없이 증식하였으나 stationary phase에서는 빠르게 생육활성을 잃는 것을 관찰할 수 있었다. 또한 생쥐를 이용한 동물 실험에서 메트로니다졸 내성균주는 생쥐의 위장 내에서 서식하는 능력을 상실함을 알 수 있었다. 그러나 이러한 생육 활성을 회복시켜주는 compensate템 mutation을 가진 균주를 쉽게 얻을 수 있었으며 이 균주는 생쥐에 감염시키면 위장 내에서 서식하는 능력을 회복함을 알 수 있었다.

방사선 육종 차조기와 백출 복합추출물의 항관절염 효과 (Anti-Arthritic Effect of Radiation Mutant Perilla frutescens var. crispa and Atractylodes macrophala koidz.)

  • 박미희;김철진;이진영;금창엽;김인선;진창현;지중구;김성규
    • 한국응용과학기술학회지
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    • 제37권1호
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    • pp.102-113
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    • 2020
  • 본 연구에서는 방사선 육종 차조기와 백출의 복합추출물의 항관절염 효능을 평가하였다. 방사선 육종 차조기와 백출 복합추출물이 세포에 미치는 독성을 확인하기 위해 RAW 264.7 세포에서 MTT 기법으로 세포 생존율을 평가하였다. 방사선 육종 차조기와 백출 복합추출물의 항염증 효능을 확인하기 위해 LPS로 염증을 유도한 RAW 264.7 세포에 방사선 육종 차조기와 백출 복합추출물을 5, 10, 25 ㎍/㎖ 농도로 처리한 후 ROS와 NO level, 염증성 사이토카인의 분비, 염증성 인자인 NF-κB, COX-2, iNOS 등의 발현을 측정하였다. 또한 type II collagen으로 유도한 관절염 모델 동물실험에서 방사선 육종 차조기와 백출 복합추출물을 33.5, 66, 133 mg/kg/day로 처리한 후 항관절염 효능을 확인하였다. 그 결과, 방사선 육종 차조기와 백출 복합추출물은 25 ㎍/㎖ 농도까지 세포독성이 없었으며, LPS로 유도된 RAW 264.7 세포에서 ROS 생성 및 NO의 생성을 5, 10, 25 ㎍/㎖ 농도에서 대조군 대비 유의성 있게 감소시켰으며, 사이토카인(IL-1β, IL-6, TNF-α)의 생성을 유의성 있게 억제시키고, NF-κB, COX-2, iNOS의 발현을 유의성 있게 감소시켜 세포 내에서 뛰어난 항염증 효과를 보였다. 관절염 모델 동물실험에서는 방사선 육종 차조기와 백출 복합추출물이 66.5, 133mg/kg 농도에서 관절염을 유의성 있게 억제시키는 효능을 나타내었다. 본 연구는 방사선 육종 차조기와 백출 복합추출물이 뛰어난 항염증 효능을 나타내는 것을 제시하며, 관절염 질환을 개선하기 위한 건강기능식품 및 치료제의 원료로 개발될 수 있다는 것을 제시한다.