• Title/Summary/Keyword: polyacrylamide gel electrophoresis

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Biochemical Studies on the Major hemolymph Proteins (MHPs) of the Silkworm, Bombyx mori (누에 체액주단백질에 관한 생화학적 연구)

  • 성수일
    • Journal of Sericultural and Entomological Science
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    • v.28 no.1
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    • pp.30-36
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    • 1986
  • Polyacrylamide gel electrophoresis and autoradiography were applied to investigate the developmental profiles of the major hemplymph proteins (MHPs) and their biosynthesis. In addition, some biochemical methods were also used to isolate and purify the MHPs. The obtained results are summarized as follows. 1. MHP-a began to appear from the 2nd day of the fourth-instar larva while MHP-b and -c were detected first on the 1st day of the fifth-instar larva. All these proteins, however, showed a drastic increase in concentration at the 2nd day of the fifth-instar larva. 2. MHP-b and -c were synthesized in fat body on early day of the fifth-instar larva, but the possibility of MHP-a synthesis in fat body was excluded. 3. MHP-b was isolated and purified by heat-treatment (6$0^{\circ}C$), gel filtration on Sephadex G-100 and column chromatography on DEAE-cellulose and CM-cellulose. Purified MHP-b showed a single band on polyacrylamide gel- and SDS-polyacrylamide gel electrophoresis.

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Tumour Specific and Immunosuppressive Components in Soluble Cell Extracts from Ovine Squamous Cell Carcinoma (면양(緬羊) Squamous Cell Carcinoma의 세포추출액(細胞抽出液)중에 함유된 종양특이(腫瘍特異) 및 면역기능저하물질(免疫機能低下物質))

  • Jun, Moo Hyung
    • Korean Journal of Agricultural Science
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    • v.13 no.2
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    • pp.299-310
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    • 1986
  • The ovine squamous cell carcinoma (OSCC)-specific and immunosuppressive properties of OSCC extracts were investigated by using the techniques of lymphocyte blastogenicity, acid dissociation-ultrafiltration and gradient polyacrylamide gel electrophoresis. It was found that OSCC extracts contained two major and one minor protein peaks by Sephadex gel fractionation. Two major peaks bear substantial amount of immunoglobulins, antigen-antibody complex and OSCC-specific fractions, and the minor peak includes immunosuppressive materials. OSCC-specific components were detected at the molecular weights of 10,000 to 100,000 daltons in the major peaks and immunosuppressive materials at the fractions with the molecular weight of 10,000 to 100,000 and < 10,000 daltons in the minor peak. When the fractions were further separated by gradient polyacrylamide gel electrophoresis, the OSCC-specific antigens were found in the slice number 4 to 6 in fraction III, and immunosuppressive materials, in the slice numbers 9 to II in fraction V. The present results were considered to provide a basis for preparation and purification of OSCC-specific and immunosuppressive materials from the crude OSCC extracts.

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Purification of $\beta$-glucanase from Bacillus subtilis Using Chromogenic Substrate (색소기질을 이용한 Bacillus subtilis의 $\beta$-glucanase 정제)

  • 이성택;양진오;정안식
    • Korean Journal of Microbiology
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    • v.26 no.3
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    • pp.223-229
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    • 1988
  • Bacillus subtilis K-4-3, which produces considerable amount of $\beta$-glucanase was selected among extracellular $\beta$-glucanase-producing bacteria isolated from soil. $\beta$-glucanase was purified by ammonium sulfate fractionation, Sephadex G-100 gel filtration and DEAE-sephacel ion exchange chromatography. The purified enzyme revealed a single band by polyacrylamide gel electrophoresis and SDS-polyacrylamide gel electrophoresis. Its molecular weight was estimated to be 17000 dalton by SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature of the purified $\beta$-glucanase were 7.0 and $50^{\circ}C$, respectively. The enzyme was strongly inhibited by 1.0mM of $Fe^{3+}$, and activated by 1.0mm of $Li^{}47+$. The absence of glucose after thin layer chromatography of reaction products revealed that the purified enzyme contains no cellobiase or laminarinbiase activity. The loberation of ki, tri-and tetra-saccharide as reaction products can be explained by endoaction of the enzyme.

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Polyacrylamide Gel Electrophoresis of the Cell Proteins from Differentiating Aspergillus niger (검정곰팡이의 분화(分化)에 있어서 폴리아크릴아마이드 겔 전기영동법(電氣泳動法)에 의한 단백질 패턴 연구(硏究))

  • Park, Yong-Ja;Kim, Jong-Hyup
    • The Korean Journal of Mycology
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    • v.8 no.3
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    • pp.159-166
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    • 1980
  • Aspergillus niger van Tieghem was cultured by the method of submerged and synchronized culture for the study of differentiation. Acid-phenol soluble cell proteins of the fungus were extracted from four stages during development. Those acid-phenol soluble proteins were separated by polyacrylamide gel electrophoresis to determine the protein patterns. A new protein band was observed from the pre-sporulation body, color density of the stained protein bands in four tubes differed according to the differentiation stages. The number of protein bands in 10% gels varied from 18 to 16, 17, and 19 according to the course of spore germination stage, conidiophore stage, phialide maturation stage and sporulation stage.

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Purification of Internal Invertase in Rhodosporidium toruloides Mating Type A Cells (Rhodosporidium toruloides의 접합형 A 세포내 Invertase의 정제)

  • Jeong, Yong-Kee;Ryu, Beung-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.6
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    • pp.725-730
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    • 1992
  • The internal invertase was purified from cell free extract of Rhodosporidium toruloides IFO 0559-M-919 by acid precipitation, ion-exchange chromatography and gel filtration to the unique enzyme protein on disc electrophoresis. We have found out that molecular weight of purified internal invertase was 90,000 by gel filtration and the purified enzyme was protein with 4 homogeneous subunits appearing as single band of 22,000daltons on SDS-polyacrylamide gel electrophoresis.

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Methodology of Genetic Engineering (遺傳工學의 方法論)

  • 이평우;윤경하;이세영
    • Korean Journal of Microbiology
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    • v.13 no.2
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    • pp.71-80
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    • 1975
  • The neurotoxin of Clostridium botulinum type B was purified from a liquid culture. The purification steps consist of ammonium sulfate precipitation of whole culture, treatment of Polymin P(0.15%, v/v), gel filtration on Sephadex G-100 at pH5.6 and DEAE-Sephadex charomatography at pH8.0. The procedure recovered 17% of the toxin assayed in the starting culture. The toxin was homogeneous by sodium dodecyl sulfate(SDS)-polyacrylamide gel electrophoresis and had a molecular weight of 163,000. Subunits of 106,000 and 56,000 molecular weight were found when purified toxin was treated with a disulfide-reducing agent and electro phoresed on SDS-polyacrylamide gels.

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Subcellular Distribution of Arginase in Leaves of Canavalia lineata (해녀콩(Canavalia lineata) 잎에서 Arginase 활성의 세포내 분포)

  • 유경희
    • Journal of Plant Biology
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    • v.33 no.1
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    • pp.73-80
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    • 1990
  • Subcellular distribution of arginase activity was measured in leaves of Canavalia lineata. Both mitochondrial and cytosolic fraction were found to contain the arginase activity. It was noticible that cytosolic fraction contained a substantial amount of arginase activity. Different mobility of arginase from these two fractions was showed on DEAE-Sephacel chromatography and polyacrylamide gel electrophoresis. Also different pI value was showed 6.3 in cytosolic and 6.7, 7.1 in mitochondiral fraction on IEF gel electrophoresis. However, canavaine-dependent-activity (CDA) of arginase in these two fractions were not different. These results indicate that heterogenity of arginase occurs in leaves of C. lineata.

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Studies on Electrophoretic Pattern and Amino Acids of Buckwheat Protein (메밀 단백질의 전기영동유형 및 아미노산 조성에 관한 연구)

  • 이미숙;손경희
    • Korean journal of food and cookery science
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    • v.8 no.4
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    • pp.379-385
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    • 1992
  • 1. Amino acid compositions were determined by amino acid analyzer. Through the analysis of these samples, it was found that glutamic acid was the most abundant; glycine, aspartic acid, lysine and threonine were rich; and tryptophan and methionine were the limiting amino acid. 2. Albumins, globulins, gliadins and glutelins were extracted from the Kangwon hull, Kangwon rice buckwheat, and wheat. The relative proportions of protein fractions were 52.45 : 10.14 : 16.61 : 20.80% in Kangwon hull buckwheat, 21.10 : 13.80 : 28.40 : 36.70% in Kangwon rice buckwheat and 6.87 : 1.65 : 42.85 : 48.6% in wheat, in the order of albumins, globulins, gliadins and glutelins. 3. Polyacrylamide gel electrophoresis (PAGE) and SDS-polyacrylamide gel electrophoresis (SDS-PAGE) were performed to identify the subfractions of each protein fraction. The electrophoregrams of PAGE showed that the same fractions of both Kangwon hull buckwheat protein and Kangwon rice buckwheat protein had very similar electrophoretic patterns to each other respectively, but there were significant differences in the patterns between buckwheat proteins and wheat proteins.

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Effect of Polyamines on purified DNA Methyltransferase from Chlamydomonas reinhardtii (Chlamydomonas reinhardtii로부터 분리, 정제된 DNA Methyltransferase 활성에 대한 Polyamine의 영향)

  • 이명민
    • Journal of Plant Biology
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    • v.32 no.4
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    • pp.331-341
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    • 1989
  • DNA methyltransferase was purified 282.6-fold from Chlamydomonas reinhardtii 21gr (mt+) gametic cell to examine the effect of polyamine on the enzyme acctivity. Polyacrylamide gel electrophoresis(PAGE) revealed at least three bands(1 major band, 2 minor bands). Among these, the major band represents DNA methyltransferase. Polyacrylamide gel electrophoresis in the presence of 0.1% sodium dodecylsulfate(SDS-PAGE) revealed a major band with M.W. 60,000. DNA methyltransferase activity was inhibited more effectively by spermine than by spermidine, and the inhibition by putrescine was smaller than spermine and spermidine. DNA methyltransferase activity was inhibited by 40% and 53% at 5mM and 20mM spermine, respectively. In the case of spermidine, the inhibition was 35% at 10mM and 44% at 20mM. However, the inhibition by putrescine appeared only above 5mM and reached about 25% at 20mM.

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Polyacrylamide Gel Electrophoresis on Ginseng Proteins (인삼 단백질분획에 대한 폴리아크릴아미드 전기영동)

  • 김춘미;황정주
    • YAKHAK HOEJI
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    • v.30 no.6
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    • pp.343-347
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    • 1986
  • Korean ginseng was purified to obtain radioprotective protein fractions by buffer extraction, ammonium sulfate fractionation, CM-cellulose column chromatography, heat inactivation and Sephadex G-75 column chromatography. The final three fractions, GI, GII and GIII were subjected to Disc-polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE. The molecular weights(M.W.) of native and denatured proteins were estimated by using regression line equations obtained from the mobilities of standard proteins. As the results, in Disc-PAGE, the GI fraction showed two protein bands with M.W. of above 213, 000 and 55, 000, GII showed one band with M.W. of 44, 000 and GIII, also one band with M.W. of 19, 000. In SDS-PAGE, GI fraction gave four subunit bands with M.W. of above 114, 000, 27, 000, 24, 000 and 19, 000, GII gave two bands with M.W. of 46, 000 and 22, 000, and GIII, one band of 19, 000.

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