• 제목/요약/키워드: Sephadex

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Sephadex G-15 또는 G-50 chromatography를 이용한 방사성 동위원소로 표지된 RNA의 정제 (Purification of Radiolabeled RNA Using Sephadex G-15 or G-50 Chromatography)

  • 유병규;이종석
    • 대한방사선기술학회지:방사선기술과학
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    • 제21권1호
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    • pp.65-68
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    • 1998
  • We attempted to purify radiolabeled RNA using Sephadex G-15 and G-50 chromatography instead of commercial RNA purification kit. In the Sephadex G-15 chromatography the major portion of RNA was eluted in the fractions ranging from 3rd to 5th whereas broad elution profile of RNA was obtained from the Sephadex G-50 chromatography. The elution profile and purity of RNA obtained from Sephadex G-15 chromatography was very similar to that by commercial RNA purification kit. Furthermore, operating time required for purification of RNA by Sephadex G-15 was rather smaller than that by commercial kit. Overall results suggest that the purification of radiolabeled RNA using Sephadex G-15 is more money and time saying than using commercial RNA purification kit.

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Rhodotorula glutinis K-24가 생산하는 Invertase Isozymes군에 관한 비교 연구 (Comparative Studies of Invertase Isozymes Produced by Rhodotorula glutinis K-24)

  • Lee, Tae-Ho;Kim, Chul;Lee, Sang-Ok
    • 한국미생물·생명공학회지
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    • 제17권4호
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    • pp.313-320
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    • 1989
  • 세포내와 세포벽에 invertase가 존재하며 catabolite inhibition을 받지않고 세포외로 구성적으로 invertase를 생산하는 Rh. glutinis K-24를 공시균주로 선택하여 세포내와 세포벽 invertase를 정제한 후 정제효소의 효소화학적 성질을 밝혔다. 세포내 invertase는 DEAE-Sephadex A-50에 의한 Batchment, DEAE-Sephadex A-50 column chromatography, gel filtration on Sephadex G-200, isoelectric focusing 등의 조작에 의해 Disc 전기영동상 단일한 효소단백질까지 정제되었다. 세포벽 invertase는 B. pumilis에 의해 생산되는 세포벽 용해효소를 작용시켜 invertase를 용출 시킨 다음 DEAE-Sephadex A-50 column chromatography, gel filtration on Sephadex G-100의 조작에 의해 부분 정제하였다. 세포내와 세포벽 Invertase의 분자량은 각각 약 310,000과 67,000이었고 세포내 경우는 subunit 분자량이 약 70,000이었다. 세포내와 세포벽 효소의 반응 최적 pH와 온도는 모두 4.0와 6$0^{\circ}C$였으며 온도 안정성도 7$0^{\circ}C$까지 거의 비슷하게 나타났다. 그러나 Km값은 세포내 효소의 경우는 4.3x$10^{-3}$M이었고 세포벽 효소의 경우는 2.1x$10^{-2}$M이었다. 그외 Inhibitor에 대한 효소의 저해양식도 양효소에 비슷한 결과를 나타내었다.

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식염수-Sephadex가 $^{99m}TC$-Sephadex의 폐분포에 미치는 영향 (Effect of Saline-Sephadex on the Organ Distribution of $^{99m}TC$-Sephadex)

  • 성호경;강신구;고주환;문광남;이장규
    • 대한핵의학회지
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    • 제5권2호
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    • pp.57-64
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    • 1971
  • The organ distribution study and the whole body scan were done in the albino rats at intervals of 5, 30, 60, 90 and 120 minutes after the intravenous injection of $^{99m}TC$-pertechnetate absorbed in Sephadex beads of $20{\sim}80$ micra in diameter. Effect of additional injection of physiological saline and saline absorbed in Sephadex beads of $20{\sim}80$ micra in diameter on the scan and organ distribution were also studied. The results were as follows. 1. Five minutes after the injection of $^{99m}TC$-pertechnetate absorbed in Sephadex beads of $20{\sim}80$ micra in diameter, Sephadex was well trapped in the lungs, with which the excellent lung scan was obtained. Two hours after the injection, kidneys were well visualized instead of lungs, which suggested that kidney acts as the excretory organ. Five minutes prior to scan, additional injection of physiological saline absorbed in Sephadex above mentioned was done. The bladder was also well visualized together with the kidneys. 2. In the distribution studies, most of radioactivity was detected in the lungs at 5 minutes and was gradually transferred chiefly to the kidneys and bladder and partly to the liver. 3. Additional injection of physiological saline resulted in a rapid transfer of $^{99m}TC$ trapped in the lung to both the kidneys and liver. 4. Additional injection of physiological saline absorbed in Sephadex beads of $20{\sim}80$ micra in diameter resulted in a rapid transfer of $^{99m}TC$ trapped in the lung to only the urinary system. 5. Results of these studies suggested that; a) Other nutrients and therapeutic compounds may be carried into the lungs along with Sephadex beads and then released in high concentration, which would exert greater therapeutic effect locally than that of the usual administration. b) Some radionuclides absorbed in Sephadex could be used as the lung scan agents, the flushing out of which by Sephadex-saline also give satisfactory renal and bladder scans. c) Other potent therapeutic radionuclides could be retained for some time by this method, which can be in the lungs easily flushed out within 2 hours.

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개 희석 정액의 다양한 filtration 처리 후 정자평가 (Evaluation of Extended Canine Semen after Different Filtration Treatment)

  • 김용준;김진영;김수희;이영준
    • 한국임상수의학회지
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    • 제24권4호
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    • pp.577-583
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    • 2007
  • It is important to obtain semen with good quality for efficient fertilization and pregnancy. To obtain these semen, various methods have been developed but most of these methods are time consuming and require costly equipment. Therefore, the objective of this research is to investigate the usability of column filtration system as quick and simple method to get sperm with better quality. Ejaculates were obtained from 5 dogs and analyzed with basic quality parameters before each filtration. Sperm concentration was adjusted to $5{\times}10^7/ml$ after dilution. The experimental groups were divided into non-filtered group(control) and filtered groups(glass wool, Sephadex 5% and Sephadex 20%). Ejaculates were filtered through each filter system and assessed by recovery rate of sperm, motility, normal morphology, CFDA/PI stain and plasma membrane integrity(hypo-osmotic swelling test, HOST). The lowest recovery rate of spermatozoa was recorded in glass wool filtration group, followed by 20% Sephadex filtration group(p<0.05). There was no significant difference between control(non-filtered) and 5% Sephadex filtration poop. Also, there was no significant difference of sperm motility assessed under light microscope among experimental groups. Morphological normality of canine spermatozoa was the highest in the glass wool filtration group and the lowest in the 5% Sephadex filtration group with no significant differences versus 20% Sephadex filtration and control group, respectively(p<0.05). Viability of canine sperm assessed by CFCA/PI staining was the highest in the glass wool filtration poop with no significant difference versus the control group, and the lowest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, respectively(p<0.05). HOS values of canine sperm was the highest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, and the lowest in the control poop with no significant difference versus glass wool filtration group, respectively(p<0.05). Therefore, these results indicated that filtration treatment for extended canine sperm would be useful method to get sperm with better quality by trapping the damaged sperm, consequently filter would be physical barrier against injured or immotile sperm.

Streptomyces속 균주로 부터 생산되는 Chitinase의 정제 및 그 성질 (Purification and Properties of an Extracellular Chitinase from Streptomyces sp.)

  • Hong, Yong-Ki;Seu, Jung-Hwn
    • 한국미생물·생명공학회지
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    • 제7권3호
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    • pp.149-155
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    • 1979
  • 자연계에서 진균류와 절족동물의 외피를 이루는 주된 다당류인 chitin(N-acetyl glucosamine polymer)의 $\beta$-1, 4-lingkage를 가수분해하는 Strepto-myces sp. 115-5 균주로부터 생성되는 chitinase를 정제하여 그 성질을 조사하였다. 48시간 진탕배양하여 생성된 chitinase를 ammonium sulfate처리, 1차 Sephadex G-100, DEAE-Cellulose, 2차 Sephadex G-100 column chromatography하여 정제하였으며 그 순도를 CM-Sephadex C-50 column chromatography 및 polyacryla-mide gel electrophoresis로서 확인하였다. 이 chitinase는 chitin과 chitosan을 가수분해 할수 있었으나 cellulose는 분해할수 없었고 chitin을 기질로서 사용하였을 경우 Km value가 3.6mg/ml이며 Vmax가 100 $\mu$mole/hr였다. Activation energy는 산 가수분해보다 훨씬 낮은 3.66kca1/mole이었고 분자량은 Sephadex G-100을 사용한 column chromatography에서 56,000 daltons으로 나타났으며, 이 chitinase의 등전점은 pH3.0에서 보여졌다.

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培養 鷄胚 筋原細胞의 融合에 미치는 鷄胚 抽出液 分劃의 영향 (The Effects of Fractions of Chick Embryo Extract on the Fusion of Cultured Chick Embryonic Myoblasts)

  • Ha, Doo-Bong;Lee, Chung-Choo;Park, Young-Chul;Lim, Woon-Ki;Yoo, Byoung-Je
    • 한국동물학회지
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    • 제28권3호
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    • pp.179-193
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    • 1985
  • 鷄胚의 筋原細胞의 증식과 融合에 미치는 계배 抽出液의 영향을 조사하였다. 1. 배양액내의 계배 추출액의 농도가 높을수록 근원세포의 증식은 촉진되었으며, 반면에 근원세포 융합은 지연되었다. 2. 계배 추출액의 단백질을 Sephadex G-75로 分劃하고, 각 분획을 근원세포의 배양액에 첨가한 결과 分子量 40,000과 22,000 dalton 사이의 분획이 근원세포의 증식과 융합을 촉진시켰다. 3. 계배 추출액의 단백질을 ammonium sulfate로 분획시켜 각 분획을 근원세포의 배양액에 첨가한 결과 70% 이상 포화 용액에서 침전하는 분획이 근원세포의 증식과 융합을 현저히 증가시켰다. 이 유효 분획을 Sephadex G-75로 재차 분획하여 각 분획의 효과를 조사한 결과 근원세포의 증식과 융합을 촉진시키는 분획이 계배 추출액을 Sephadex G-75로 분획하여 얻은 유효 분획과 거의 동일한 효과를 나타내었다.

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Quality and Fertility of Post Thaw Sephadex Filtered Bull Semen

  • Kumar, A.;Singh, J.;Nanda, A.S.;Pangaonkar, G.R.
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권6호
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    • pp.755-759
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    • 2004
  • The present investigation was carried out to assess the effect of Sephadex (G-15) filtration on the post thaw bull semen quality and conception rate. Post thaw unfiltered (control) and Sephadex filtered semen from four healthy bulls (three cross bred and one pure bred Holstein Friesian) were subjected to microscopic examination viz. sperm concentration, individual motility, live sperm count and sperm morphology. Sixty-two healthy, normal cycling crossbred cows were inseminated with post thaw unfiltered (n=32) and filtered semen (n=30). Sephadex filtration of post thaw semen significantly (p<0.05) decreased total sperm concentration and sperm with abnormal head, mid piece and tail. The overall average total sperm concentration, head and tail defects in filtered semen decreased significantly (53.4, 1.2 and 6.4 million) than in the unfiltered semen (80.4, 2.4 and 15.7 million, respectively). However, after filtration significant (p<0.05) increase in overall average motile and live sperm concentration were observed (38.8 and 38.0) as compared to unfiltered semen (29.2 and 32.0 million, respectively). The overall conception rate recorded was 21.9% with post thaw unfiltered semen and 56.7% with filtered semen. It was concluded that Sephadex filtration of post thaw semen improved its quality and conception rate.

Aspergillus niger가 생산하는 섬유소 분해효소의 기질에 대한 특이성 (Substrate Specificity of Cellulase from Aspergillus niger)

  • 오태광;박관화;신현경;김재욱
    • Applied Biological Chemistry
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    • 제28권3호
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    • pp.162-166
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    • 1985
  • Aspergillus niger가 생산하는 섬유소 분해 효소를 Sephadex G-150, DEAE-Sephadex 및 Sephadex G-75를 통해서 세개의 C.M.C 분해 효소와 2개의 고결정형(高結晶形) 섬유소 분해능을 가진 효소인 ${\beta}-1,4-D-cellobiohydrolase$로 분리하고 기질에 대한 특이성을 연구하였다. 효소의 기질에 대한 분해 특성 및 기질의 X-선 회절결과로 ${\beta}-1,4-D-cellobiohydrolase$는 무정형 섬유소에 대한 분해력은 거의 없고 주로 결정형 섬유소에 특이성이 컸다. Cx 효소와 ${\beta}-1,4-D-cellobiohydrolase$가 동시에 작용할 때는 상승효과를 보였다.

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