• 제목/요약/키워드: P10 partial protein

검색결과 161건 처리시간 0.026초

벼검은줄오갈병바이러스 외피단백질 유전자 단백질 발현과 항혈청 제작 (In Vitro Expression and Antibody Preparation of Rice black-streaked dwarf virus Coat Protein Gene)

  • 이봉춘;조상윤;배주영;김상민;신동범;김선림
    • 식물병연구
    • /
    • 제22권1호
    • /
    • pp.32-37
    • /
    • 2016
  • 본 연구에서는 RBSDV의 외피단백질 P10을 코드하는 S10을 E. coli에서 발현시켰다. RBSDV-miryang isolate (GenBank JX994211)로부터 추출한 게놈 dsRNA을 주형으로 S10의 특이적인 primer를 사용하여 P10의 N-말단영역(1-834 nt, 1-278 aa)을 RT-PCR에 의해 증폭하였다. 증폭된 RBSDV S10-N (1-834 nt)을 발현 벡터 pET32a(+)에 클로닝하여 E. coli BL21(DE3)에서 발현시킨 후 Ni-NTA affinity column으로 발현된 단백질을 정제하였다. 정제된 단백질을 면역 동물에 주사하여 항혈청을 제작하였다. 제작된 항혈청은 Western blot 및 ELISA 분석으로 RBSDV와의 특이성을 확인하였다. 본 연구에서 RBSDV 한국 isolate의 항혈청이 제작되었으며 금후 혈청학적 연구의 좋은 재료로 활용될 수 있을 것으로 기대한다.

Construction of a Novel Recombinant Bombyx mori Nuclear Polyhedrosis Virus Producing the Fluorescent Polyhedra

  • Kang, Seok-Woo;Yun, Eun-Young;Woo, Soo-Dong;Goo, Tae-Won;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • 제3권1호
    • /
    • pp.75-81
    • /
    • 2001
  • We have constructed a novel recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV) producing the green fluorescent polyhedra. For the production of the fluorescent polyhedra, partial polyhedrin gene containing KRKK as nuclear localization site from the BmNPV polyhedrin gene and the green fluorescent protein (gfp) gene were introduced under the control of p10 promoter of BmNPV. The recombinant BmNPV was stably produced fluorescent polyhedra in the infected Bm5 cells and the morphology of the fluorescent polyhedra was similar to that of wild-type BmNPV. The fluorescent polyhedra had 32 kDa native polyhedrin and 41 kDa fusion protein. From these data, we have further developed a novel BmNPV p10-based transfer vector producing recombinant polyhedra with foreign gene Product. The novel BmNPV P10-based transfer vector is composed of partial polyhedrin gene, factor Xa, and multiple cloning sites.

  • PDF

Identification and Characterization of pH-Regulated Genes in Saccharomyces cerevisiae

  • Hong, Sung-Ki;Choi, Eui-Yul
    • Journal of Microbiology
    • /
    • 제34권4호
    • /
    • pp.327-333
    • /
    • 1996
  • Yeast, like many other microbes, encounters large variations in ambient pH in their natural environments. Microorganisms capable of growing over a wide pH range require a versatile, efficient pH homeostatic mechanism protecting intracellular processes against extremes of pH. In several organisms, fusions to the bacterial lacZ gene have been extremely useful for the identification of genes expressed at different time during the life cycle or under different growth conditions. In this study, using the lacZ gene screening system, we surveyed a large number of yeast strains with lacZ insertion to identify genes regulated by pH. A yeast genomic library was constructed and inserted with lacZ by a shuttle mutagenesis procedure. The yeast transformants were individually picked up with a toothpick, replica-plated, and grown in alkaline pH medium. Among the 35,000 colonies screened, 10 candidate strains were identified initially by the $\beta$-gal assay. We finally confirmed two yeast strains carrying the genes whose expression are strictly dependent on pH of growth medium. One of the fusions showing a 10-fold induction in expression level in response to alkali pH was selected and further characterized. The pH-regulated gene was cloned by inverse PCR and a partial sequence of the gene was determined. Identification and characterization of the gene is currently under investigation.

  • PDF

Partial Freezing에 의한 어육의 선도유지 효과에 대하여 1. Partial Freezing에 의한 숭어의 선도 및 어묵형성능의 변화 (Effect of Partial Freezing as a Means of Keeping Freshness I. Changes in Freshness and Gel Forming Ability of Mullet Muscle during Storage by Partial Freezing)

  • 이용우;박영호
    • 한국수산과학회지
    • /
    • 제18권6호
    • /
    • pp.529-537
    • /
    • 1985
  • 숭어를 시료어로 하여 이것을 빙장, $-3^{\circ}C$에서의 partial freezing 및 $-3^{\circ}C$에서의 동결저장을 하여 저장 중의 시료육의 pH, VBN 및 K 치의 변화, 지질의 POV와 TBA 치의 변화, 육단백질의 비단백질소화합물, 근형질단백질. 근원섬유단백질, 알칼리가용단백질 및 기질단백질 등의 조성변화를 분석하여 partial freezing에 의한 선도유지효과를 검토하고, 아울러 저장 기간별로 어묵을 제조하여 그 겔강도, 압출수분율, 절곡강도 및 texture 등을 측정, 비교하므로서 시료육의 어묵형성능의 유지효과도 검토하였다. 1. 시료어의 저장 중 pH가 최저치에 달하는 시간은 동결저장, partial freezing, 빙장의 순으로 길었다. VBN의 변화에 있어서는 동결저장과 partial freezing의 경우는 큰 변화를 나타내지 않았으나, 빙장의 경우는 그 증가 속도가 비교적 빨랐다. 2. 저장 중의 K 치의 변화에 있어서 빙장 및 Partial freezing의 경우는 급속히 증가하였으나, 그 증가폭은 partial freezing의 경우가 적어서 저장 6일째에 빙장의 경우는 약 $50\%$, partial freezing의 경우는 약 $30\%$를 나타내었다. 그러나, 동결저장의 경우는 거의 증가하지 않았다. 3. TBA 치의 변화에 있어서 빙장의 경우는 서서히 증가하여 저장 15일경에 최고치에 달한 후 감소하는 변화를 보였으나, partial freezing 및 동결저장의 경우는 큰 변화를 나타내지 않았다. POV의 변화에 있어서 빙장의 경우는 저장 9일경까지 급속히 증가하였다가 감소하는 변화를 보였으나, partial freezing 및 동결저장의 경우는 극히 완만하게 증가하는 변화를 보였다. 4, 저장 중의 단백질조성의 변화에 있어서 비단백질소화합물, 근형질단백질 및 기질단백질은 큰 변화가 없는데 비하여, 근원섬유단백질은 감소하고, 알칼리가용단백질은 증가하는 변화를 보였다, 저장기간에 따라 근원섬유단백질이 감소하는 정도와 알칼리가용단백질이 증가하는 정도는 빙장, partial freezing, 동결저장의 순으로 컸다. 5. 시료어의 저장 조건별로 제조한 어묵의 겔강도, 절곡강도 및 texture의 저장 기간에 따른 감소의 정도는 빙장의 경우가 가장 크고, 다음이 partial freezing의 경우였으며, 동결저장의 경우는 변화가 적었다. 10일간 저장한 시료어의 경우 어묵의 겔강도는 빙장을 한 것은 대조구의 것의 약 $30\%$, partial freezing을 한 것은 약 $60\%$, 동결저장을 한 것은 약 $97\%$를 나타내었다. 어묵의 압출수분율은 원료어의 저장 기간에 따라 증가하였는데, 15일간 저장한 시료어의 경우 어묵의 압출수분율은 빙장을 한 것은 대조구의 것의 약 2.1배, partial freezing을 한 것은 약 1.5배, 동결저장을 한 것은 악 1.1배를 나타내었다.

  • PDF

배합사료의 어분 대체원으로 막걸리부산물 첨가가 조피볼락 치어의 성장에 미치는 영향 (Effects of Dietary Inclusion of Distillers Dried Grain as a Partial Replacement for Fish Meal on Growth Performance of Juvenile Rockfish Sebastes schlegeli)

  • 배기민;이상민
    • 수산해양교육연구
    • /
    • 제27권2호
    • /
    • pp.390-398
    • /
    • 2015
  • A feeding experiment was conducted to determine the use of distillers dried grain (DDG) as a partial replacement for fish meal in the diet for juvenile rockfish, Sebastes schlegeli. Four iso-nitrogenous (50% crude protein) and iso-caloric (4.3 kcal/g) diets (designated as DDG0, DDG7, DDG14, and DDG21) were formulated to contain 0, 7, 14, and 21% DDG. Triplicate groups of juvenile rockfish (initial body weight, $10.2{\pm}0.2g$) were fed one of the experimental diets to visual satiety twice a day (09:00 and 17:00) for 8 weeks. At the end of the feeding trial, survival of rockfish was above 97% and not affected by dietary DDG levels (P>0.05). Weight gain, feed efficiency and daily feed intake of juvenile rockfish were significantly decreased with increase of dietary DDG levels (P<0.05). Condition factor, hepatosomatic index and visceralsomatic index of juvenile rockfish were not significantly affected by dietary DDG levels (P>0.05). No significant differences were observed in the contents of moisture, crude protein, crude lipid and ash of the whole body and dorsal muscle in juvenile rockfish fed the experimental diets (P>0.05). Therefore dietary inclusion of DDG as a replacement for fish meal could depress the growth of juvenile rockfish.

참전복 치패용 배합사료의 단백질원로서의 밀배아박 첨가효소 (Utilization of Wheat Germ Meal As a Protein Source of Formulated Diet for Juvenile Abalone (Haliotis discus hannai))

  • 이상민;이종관;이종하;임영수
    • 한국양식학회지
    • /
    • 제12권4호
    • /
    • pp.317-322
    • /
    • 1999
  • A growth trial was conducted to investigate the utilization of wheat germ meal as a protein source of formulated diet for juvenile abalone (Haliotis discus hannai). Four replicate groups of the abalone average weighing 150mg were fed one of four isonitrogenous (33%) and isolipidic (6%) diets containing 0%, 10%, 20%, or 30% wheat germ meal for 18 weeks. In addition, these formulated diets were compared with commercial diet. Survival rate, weight gain, soft body weight , and shell growth of abalone fed diets containing 0%, 10%, 20%, or 30% wheat germ meal were not different (P>0.05) from those of abalonn fed the control diet and commercial diet. There were no significant differences (P>0.05) in soft body composition of moisture, protein and lipid. It si concluded that wheat germ meal were be used as a partial protein source of formulated diet for juvenile abalone.

  • PDF

The Use of Meat Meal as a Dietary Protein Source Replacing Fish Meal in Juvenile Rockfish Sebastes schlegeli

  • Lee, Yong-Whan;Lee, Sang-Min
    • 한국양식학회지
    • /
    • 제18권2호
    • /
    • pp.92-97
    • /
    • 2005
  • This study examined the partial replacement of the fish meal with meat meal in practical diets for juvenile rock-fish. Five isonitrogenous (48% CP) diets were prepared to contain meat meal at 0% (control), 10%, 20%, 30% and 40% with substituting the mackerel meal in the control diet. Three replicate groups of fish (initial average weight, 4.1g) were hand-fed to visual satiety two times daily for 8 weeks. Survival (>93%) and daily feed intake were not significantly different (P>0.05) among treatments. The best weight gain, feed efficiency and protein efficiency ratio were obtained from fish fed the diets containing 0% and 10% meat meal, and were not significantly different (P>0.05) to those of fish 134 diet containing 20% meat meal. Condition factor, visceralsomatic index and hepatosomatic index were not influenced by dietary meat meal levels. The contents of crude protein and ash of whole body were not significantly affected (P>0.05) by dietary meat meal levels, whereas crude lipid content of fish fed the diets containing 30% and 40% was lower than that of fish fed the control diet. Proximate composition of liver was not influenced by dietary meat meal level (P>0.05). The data obtained in this study indicate that a diet containing $10{\sim}20%$ meat meal could be used for least-cost formulation in juvenile rockfish diet.

The Establishment of Tumor Necrosis Factor Receptor-associated Protein1 (TRAP1) Transgenic Mice and Severe Fat Accumulation in the Liver of TRAP1 Mice during Liver Regeneration

  • Im, Chang-Nim;Zheng, Ying;Kim, Sun Hye;Huang, Tai-Qin;Cho, Du-Hyong;Seo, Jeong-Sun
    • Interdisciplinary Bio Central
    • /
    • 제5권4호
    • /
    • pp.9.1-9.7
    • /
    • 2013
  • Introduction: Tumor necrosis factor receptor-associated protein 1 (TRAP1) is a mitochondrial heat shock protein (HSP), which belongs to HSP90 family. It plays important roles in regulating mitochondrial integrity, protecting against oxidative stress, and inhibiting cell death. Recent studies suggest that TRAP1 is linked to mitochondria and its metabolism. In this study, we established TRAP1 transgenic mice and performed partial hepatectomy (PH) on wild-type (WT) and TRAP1 transgenic mice to investigate the function of TRAP1 during liver regeneration. Results and Discussion: We found that TRAP1 was highly expressed in liver as well as kidney. In addition, liver regeneration slightly decreased together with increased fatty liver and inflammation at 72 hr after PH in TRAP1 transgenic mice compared with WT control group mice. Concomitantly, we observed decreased levels of p38 protein in TRAP1 transgenic mice compared with WT control group mice. These results suggest that TRAP1 plays a critical role in liver energy balance by regulating lipid accumulation during liver regeneration. Conclusions and Prospects: To our knowledge, we reported, for the first time, that liver regeneration slightly reduced together with increased fat accumulations after PH in TRAP1 transgenic mice compared with WT control group mice. Concomitantly, we observed decreased levels of p38 protein in TRAP1 transgenic mice compared with WT control group mice. Overexpression of TRAP1 might affect liver regeneration via disturbing mitochondrial function leading to fatty liver in vivo.

잿빛곰팜이병균 Botrytis cinera가 분비하는 Polygalacturonase의 부분정제와 특성 (Partial Purification and Properties of Polygalacturonase Produced by Botrytis cinerea)

  • 나유진;김재원;정영륜;허남응;조광연
    • 한국식물병리학회지
    • /
    • 제10권3호
    • /
    • pp.215-221
    • /
    • 1994
  • Polygalacturonase (PG) produced by Botrytis cinerea in the culture broth containing citrus pectin as a carbon source was partially purified and characterized. PG was produced on a range of carbon sources such as starch, glycerol, cellobiose, and Na+-PAG with total activities of 34.8, 32.0, 29.2, 27.8 units, respectively. The specific activity was highest with 2316.7 units on Na+-PGA. Proteins of culture filtrate were concentrated with polyethylene glycol and acetone and applied to a hydroxyapatite column. Among three active fractions collected from the column, the reaction containing the highest PG activity was resolved by a Q-sepharose column. The active fraction from the Q-sepharose column was further purified by HPLC Mono Q column. The partially purified enzyme was analyzed by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Among a few protein bands revealed, the amount of the protein of which molecular weight estimated to be 43 kDa coincided with the PG activity. The partially purified PG had optimal temperatures between 35~55$^{\circ}C$ and pH between 4.5~5.5.

  • PDF