• Title/Summary/Keyword: 2-D PAGE

검색결과 322건 처리시간 0.037초

효소적 가수분해에 따른 두유와 전두유의 단백질 변화 (Protein Changes in Soymilk and Whole Soymilk due to Enzymatic Hydrolysis)

  • 장세영;신경아;박난영;방광웅;정용진
    • 한국식품저장유통학회지
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    • 제15권6호
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    • pp.903-908
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    • 2008
  • 본 연구는 두유와 전두유의 효소적 가수분해에 따른 단백질의 변화를 조사하였다. 총 유리아미노산 함량을 조사한 결과 두유(SM)과 전두유(WSM)에 비해서 저분자 두유(LSM)와 저분자 전두유(LWSM)에서 높게 나타났다. 필수아미노산 함량은 SM과 LSM에서 비슷하였으나 LWSM은 WSM보다 높게 나타났다. SDS-PAGE 전기영동 패턴 분석결과 SM과 WSM에서는 $33{\sim}72\;kDa$의 고분자가 존재하였으나 LSM와 LWSM에서는 17 kDa 이하의 저분자 단백질만 나타났다. 또한 이차원 전기영동한 결과 SM과 LSM에서의 고분자 단백질 spot이 WSM와 LWSM에서는 다양한 크기의 저분자 단백질 spot으로 변환되어 효소가수분해에 의해 고분자 단백질이 저분자화 되는 것으로 나타났으며, 향후 단백질 spot의 분리 및 특성에 관한 연구가 필요하다.

벼잎 절편에서 Polyamine이 엽록소 함량 및 Chloroplast Peroxidase활성에 미치는 영향에 관한 연구 (Studies on the Effect of Polyamine on Chlorophyll Contents and Chloroplast Peroxidase Activities in Rice Leaf Segments)

  • 표병식;김영준강영희
    • KSBB Journal
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    • 제8권2호
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    • pp.115-121
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    • 1993
  • 호르몬이 결여되고 polyamine이 농도별로 처리된 M MS배지에서 10일 동안 발아생장(light, 16hrs: d dark, 8hrs) 시켜서 얻은 벼 잎 절편에서 엽록소 함 량은 polyamine 처리구가 대조구에 비해 높았으며, s spermine(O.OlmM, O.lmM, ImM) 이 가장 효과적이였다. 한편 chloroplast peroxidase의 활성은 polyamine 처리구가 대조구에 비해 전반적으로 높았으며, 특히 1mM spermidine 처리구는 약 100% 정도 활성을 증가시켰다. SDS-PAGE에 의해 chloroplast thylakoid membrane protein의 band를 조사한 결과 polyamine 처리구와 대조구에서 56, 2 25Kd의 major band를 얻었으며 이 band들의 total a area는 polyamine 처리구가 대조구 보다 더 높았다. 이러한 결과들은 벼 유식물에서 엽록체 발달에 p polyamine이 중요한 인자로 작용한 것으로 사료된다.

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초기 임신돈에 있어서 자궁분비단백질의 전기영동상과 면역억제력에 관한 연구 (A Study on Electrophoretic Pattern and Immunosuppressive Activity of Uterine Secretory Protein in Early Pregnant Sows)

  • 김경순;정영채;김창근;한기영;김형태;윤종택;김종대;최선호
    • 한국가축번식학회지
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    • 제13권3호
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    • pp.179-187
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    • 1989
  • The experiment was carried out to study the profile of uterine specific protein during early pregnancy in sows and to test it's immunosuppressive activity. Uterine protein samples were obtained by flushing the uterine horn on Day 3, 6, 9, 12 and 15 of the estrous cycle and the pregnancy respectively and the protein concentration of each sample was determined. The change of uterine protein was examined by sodium dodecyl sulfate(SDS)-PAGE. The immunosuppressive activity of uterine secretory protein was investigated according to the lymphocyte blastogenesis response to mitogen. The results of this experiment are summarized as follows ; 1. The uterine protein during estrous cycle and early pregnancy was relatively constant up to Day 9, but increased on Day 12. Maxium total protein values were found on Day 15. The concentration of serum proteins were about 82-95 mg during estrous cycle, but decreased to about 70-82 mg during early pregnancy. 2. The proteins components similar electrophoretic patterns(PAGE) that were no differences (band ; a, b, c, d, e, f, g, I) on Days 3, 6 and 9 of the estrous cycle and pregnancy. But there were more 2 bands specifically on Day 12 of the pregnancy and on Day 15 of estrous cycle and showed more 4 bands on Day 15 of early pregnancy. They seemed to be acidoprotein and their average molecular weight were 38,000, 22,300 and 12,600. 3. When uterine protein were added 500$\mu\textrm{g}$/ml, there was no immunosuppresive activity on Day 3 of estrous cycle and lymphocyte blastogenesis was slightly suppressed on Day 3 of pregnancy. The immunosuppressive activity on Day 9 of estrous cycle and pregnancy appeared in 500$\mu\textrm{g}$/ml and 150$\mu\textrm{g}$/ml, respectively the uterine protein on Day 12 and 15 showed immunosuppresive activity, which at the level of 150$\mu\textrm{g}$/ml during non-pregnancy and at the level of 100 to 125$\mu\textrm{g}$/ml during early pregnancy, respectively.

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Biochemical Performance and Quantitative Assessment of F1 Hybrid of Two Ecoraces of Tropical Tasar Silkworm Antheraea Mylitta Drury (Lepidoptera: Saturniidae)

  • Lokesh, Gangadharaiah;Tirkey, Sushma Rani;Srivastava, Ashok Kumar;Kar, Prasant Kumar;Sinha, Manoj Kumar
    • International Journal of Industrial Entomology and Biomaterials
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    • 제26권2호
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    • pp.67-73
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    • 2013
  • Antheraea mylitta Drury is basically a crossbreeding species, as such it seems to be potentially a good material for the exploitation of heterosis. In the present study F1 hybrid of wild ecorace Laria (L) and semi-domestic Daba (D) was raised and evaluated for various quantitative traits and biochemical parameters during larval stage. Improved fecundity ($+18{\pm}1.8%$ and higher egg hatching rate ($+10.96{\pm}1.3%$) was recorded in the F1hybrid ($L{\times}D$). Biochemical parameters studied in the hemolymph, midgut and fatbody of the larva showed significantly higher (P<0.05) total proteins and carbohydrate concentration besides digestive enzyme activity. Correspondingly SDS-PAGE revealed more number of protein bands in the hemolymph sample of F1s, ranging between 29 kDa to 66 kDa compared to parental lines. The present study demonstrates the positive heterosis effect in the F1 hybrid of Laria ${\times}$ Daba. Biochemical analysis indicates that, there is possibilities of exploitation of hybrids with specific parents targeted for desirable commercial traits (silk yield and fecundity). Moreover, most of these biochemical parameters can be used as markers to analyze the genetic improvement in the tasar silkworms.

Proteomic Assessment of Dung Beetle, Copris tripartitus Immune Response

  • Suh, Hwa-Jin;Bang, Hea-Son;Kim, Seong-Ryul;Yun, Eun-Young;Park, Kwan-Ho;Kang, Bo-Ram;Kim, Ik-Soo;Jeon, Jae-Pil;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • 제17권2호
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    • pp.217-221
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    • 2008
  • Dung beetle larvae at the $3^{rd}$ instar were injected with lipopolysaccaride and inducible proteins were examined within a pI level of 3-10 and a size level by proteomics, including 1-D SDS PAGE analysis and antibacterial assay. The immune infected larvae extracts provided seven protein bands in one-dimensional electrophoresis and its antibacterial activity also checked. Hemolymph protein from immune infected larvae of the dung beetle were separated by twodimensional gel electrophoresis and compared with those from native larvae. In 2-D gel electrophoresis, we detected 63 immune infected unique and 32 up-regulated proteins, and 36 proteins that were down-regulated or not present in treated gel. Ten protein spots from unique proteins and those presented as different level of abundance in infected and native larvae were specially expressed. These differentially expressed proteins were proposed to be involved in the defense mechanism against microorganism.

Change in Levels of Endogenous Hormone and Detection of Adventitious Bud-Related Protein during Culture of Hybrid Poplar Explants

  • Song, Jae-Jin
    • Journal of Plant Biology
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    • 제38권2호
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    • pp.143-151
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    • 1995
  • Addition of plant growth hormones [0.01 mg/L NAA and 0.2mg/L benzyladenine (BA)] to a woody plant medium stimulated the adventitious bud formation of poplar explants during culture. Endogenous IAA content increased rapidly at the initial culture stage and then decreased, being followed by rapid increment again at the late culture. But the content of trans-zeatin riboside (t-ZR) increased continuously during the culture. Cytoplasmic soluble proteins were analyzed by one- and two-dimensional SDS-PAGE. Increased amount of 40 kD band was detected by one-dimensional electrophoresis using Coomassie Blue staining during the culture and two distinctive proteins whose mol wt is 40,000 were detected by two-dimensional electrophoresis using autoradiography and these proteins were synthesized continuously prior to the adventitious bud formation. When the midvein segments were transferred to the actinomycin D-containing medium, the spots of adventitious bud-related proteins(ABRPs) did not disappeared but weakened in intensity. So, it is concluded that genes coding for the ABRPs are regulated to some degree at the transcriptional level. Also, they were not observed in BA-free medium, suggesting that these proteins be regulated by cytokinin, which made then possible to form the adventitious bud.

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A Novel Ginsenosidase from an Aspergillus Strain Hydrolyzing 6-O-Multi-Glycosides of Protopanaxatriol-Type Ginsenosides, Named Ginsenosidase Type IV

  • Wang, Dong-Ming;Yu, Hong-Shan;Song, Jian-Guo;Xu, Yu-Feng;Liu, Chun-Ying;Jin, Feng-Xie
    • Journal of Microbiology and Biotechnology
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    • 제21권10호
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    • pp.1057-1063
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    • 2011
  • Herein, a novel ginsenosidase, named ginsenosidase type IV, hydrolyzing 6-O-multi-glycosides of protopanaxatriol-type ginsenosides (PPT), such as Re, R1, Rf, and Rg2, was isolated from the Aspergillus sp. 39g strain, purified, and characterized. Ginsenosidase type IV was able to hydrolyze the 6-O-${\alpha}$-L-($1{\rightarrow}2$)-rhamnoside of Re and the 6-O-${\beta}$-D-($1{\rightarrow}2$)-xyloside of R1 into ginsenoside Rg1. Subsequently, it could hydrolyze the 6-O-${\beta}$-D-glucoside of Rg1 into F1. Similarly, it was able to hydrolyze the 6-O-$_{\alpha}$-L-($1{\rightarrow}2$)-rhamnoside of Rg2 and the 6-O-${\beta}$-D-($1{\rightarrow}2$)-glucoside of Rf into Rh1, and then further hydrolyze Rh1 into its aglycone. However, ginsenosidase type IV could not hydrolyze the 3-O- or 20-O-glycosides of protopanaxadiol-type ginsenosides (PPD), such as Rb1, Rb2, Rb3, Rc, and Rd. These exhibited properties are significantly different from those of glycosidases described in Enzyme Nomenclature by the NC-IUBMB. The optimal temperature and pH for ginsenosidase type IV were $40^{\circ}C$ and 6.0, respectively. The activity of ginsenosidase type IV was slightly improved by the $Mg^{2+}$ ion, and inhibited by $Cu^{2+}$ and $Fe^{2+}$ ions. The molecular mass of the enzyme, based on SDS-PAGE, was noted as being approximately 56 kDa.

Web 3D를 이용한 3차원 가상도시공간정보 구축 (Construction of Three Dimensional Virtual City Information Using the Web 3D)

  • 유환희;조정운;이학균
    • 한국측량학회지
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    • 제20권2호
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    • pp.119-126
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    • 2002
  • 최근 Web 3D기술의 발달과 인터넷에서의 가상현실 구현에 관한 기술개발로 인하여 웹에서 3차원으로 정보를 제공하려는 연구가 활발히 진행되고 있다. 특히, 인터넷에서 가상세계를 구현하는 표준 언어인 VRML과 HTML문서에 있는 자바 애플릿과의 인터페이스를 제공하는 EAI방식을 이용하여 3차원 공간에서 도시정보를 웹 서비스하기 위한 다양한 어플리케이션 개발이 이루어지고 있다. 본 연구는 수치지도와 위성영상, VRML 등을 이용하여 3차원 가상도시를 구현하고, 웹(Web)에서 도시생활정보를 제공하는 3차원 가상도시공간정보를 EAI를 이용하여 구축함으로써 사용자 중심의 보다 현실감 있는 건물의 위치정보 및 생활정보를 제공할 수 있었다. 또한 텍스쳐링, 파노라마, 네비게이션 등 Web 3D 기술을 적용하여 다양한 경관분석과 경로탐색을 가능하게 하였고, 데이터베이스와의 연동을 통한 속성정보의 검색뿐만 아니라, ASP를 이용하여 웹에서 데이터 업로드가 가능하도록 설계함으로써 실시간으로 도시생활정보를 제공할 수 있게 구축하였다.

돌돔(Oplegnathus fasciatus) somatolactin cDNA의 분석 (Characterization of Somatolactin cDNA from Rock Bream (Oplegnathus fasciatus))

  • 강현실;여인규;이제희
    • 생명과학회지
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    • 제13권6호
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    • pp.805-813
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    • 2003
  • 돌돔 (Oplegnathus fasciatus) SL을 암호화하는 cDNA clone을 뇌하수체로부터 RT-PCR 방법에 의해 획득하였다. 돌돔 SL cDNA의 길이는 1636 bp로서 24개의 아미노산인 signal peptide와 207개 의 aa으로 구성된 mature protein을 암호화하는 696 bp의 open reading frame을 갖고 있다. 또한, 돌돔 SL 아미노산에는 이황화 결합에 관여하는 7개의 시스테인 잔기 $(Cys^{5},\; Cys^{15},\; Cys^{42},\; Cys^{65},\; Cys^{181},\; Cys^{198}\$$Cys^{206})$와 두 개의 potential N-glycosylation site인 $Asn^{121}$$Asn^{153}$을 확인하였다. 돌돔 SL은 goldfish와 channel catfish를 제외한 다른 경골어류 SL에 아미노산 서열은 61.1∼92.6%, 뉴클레오타이드 서열은 63∼92.6%의 일치를 나타낸다. 아미노산 서열 alingment에서 돌돔 SL은 다른 어류 SL에 공통적인 4개의 conserved domain $(A_{SL},\; B_{SL},\; C_{SL}$$D_{SL})$을 갖고 있음을 확인하였다. 이들중 $A_{SL},\; B_{SL}$,과 $D_{SL}$,은 경골어류 growth hormone과 prolactin에도 잘 보존되어 있었다 재조합 돌돔 SL 단백질을 E. coli에서 생산하기 위해 돌돔 SL cDNA를 발현벡터에 클로닝하여 단백질의 발현을 유도하였다 발현된 단백질은 SDS-PAGE에 의해 분자량 약 27 kDa의 재조합 단백질의 발현을 확인하였다.

Streptomyces violaceoruber 유래 Alginate Lyase의 정제 및 Sodium Alginate 가수분해 올리고당의 중합도별 Bifidobacterium spp.과 Lactobacillus spp.에 대한 생육활성 (Purification of Alginate Lyase from Streptomyces violaceoruber and the Growth Activity of Intestinal Bacteria by Degree of Polymerization of Alginate Hydrolysates)

  • 윤민;박영서;박귀근
    • 산업식품공학
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    • 제21권2호
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    • pp.103-109
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    • 2017
  • DEAE sephacel anion chromatography 및 SP sepharose cation chromatography에 의해 Streptomyces violaceoruber 유래 alginate lyase의 정제를 수행하여 비활성 14.59 units/mL 정제배율 40.64배를 나타내었다. Tricine SDS-PAGE에 의한 단일밴드를 확인하였고, 분자량은 23.3 kDa으로 결정되었다. 정제효소에 의해 sodium alginate를 가수분해하여 1차 activated carbon column chromatography와 2차 bio gel P-2 gel filtration에 의해 당가수분해물을 분리 회수하여 TLC와 FACE를 통해 중합도를 확인하고 Timell's method에 의해 hetero type M/G-oligosaccharide 중합도 6, 8로 결정되었다. B. animalis, B. bifidum, B. breve, B. infantis, B. longum와 L. acidophilus, L. casei, L. reuteri에 생육활성에 대한 중합도 6, 8의 영향을 검토하기 위하여 modified-MRS media에 탄소원으로 중합도 6, 8를 대체하여 생육활성을 비교한 결과 B. longum에서는 D.P. 6 M/G-oligosaccharide를 탄소원으로 대체한 경우 표준 MRS배지와 비교하여 4.25배, D.P. 8에서 6.44배의 상대활성을 나타내어 가장 우수한 생육활성을 나타내었으며, B. bifidum의 경우에서도 D.P. 6에서 3.27배, D.P. 8에서 5.4배의 상대활성을 나타내었다. 이외에도 B. animalis, B. breve그리고 L. casei에 있어서도 D.P. 8의 경우 3배의 상대활성을 나타내었으나, L. reuteri에 대한 D.P. 8의 경우에서는 표준 MRS media와 비교하여 0.29배로 감소하였다. 결과적으로 D.P. 8의 올리고당이 D.P. 6의 올리고당보다 생육활성에 크게 기여하는 것으로 나타났다.