• 제목/요약/키워드: isozyme

검색결과 521건 처리시간 0.034초

한국산 검정망둑(Tridentiger obscurus)과 민물검정망둑(T.brevispinis)의 분류학적 고찰

  • 김종범;양서영
    • Animal Systematics, Evolution and Diversity
    • /
    • 제11권3호
    • /
    • pp.359-377
    • /
    • 1995
  • 전국에서 채집된 검정망둑과 민물검정망둑의 지리적 변이를 조사한 결과 두 종은 반문형태상 뚜렷한 차이가 있었고 유전자 분석결과에서도 두 종은 3개의 유전적 표식인자 (Me-1, Gp-4, Aco)를 가지는 별개의 분류군으로 확인되었다. 그러나 유전적 근연치는 S=0.813 , 유전적 차이치는 D=0.192로 종 수준 이하의 유전적 분화를 보였으며두 종의 동서하천에서 미세분포 상황 및 생식적 격리 수준을 조사한 결과, 두 종은 서식처 분리가 ENfut하여 검정망둑은 주로 염수역에, 민물검정망둑은 주로 담수역에 분포하며 이들 사이에 유\ulcorner 교환이 없는 것으로 확인되었으나 서식처 분리가 깨진 여천 방죽천 집단의 경우, 두 종간에는 유전자 교환이 일어나며 F2 이상의 잡종개체를 포함하여 높은 비율(25.6%) 의 자연잡종이 발생되는 것으로 보다 두종은 교배전 격리기작 (prmatign isolating mechanism) 이 불완전한 것으로 확인되었다. 그러나 F-statistics 분석결과 $F_{IS}$(inbredding coefficient) 값은 두 종간에 자유교배가 일어나지 않음을 보여 검정망둑과 민물검정망둑은 현재 아종에서 종으로 분화중에 있는 반종(semispecies)으로 추정되며 따라서 학명은 T.obscurus obscurus(검정망둑)와 T.obscurus brevispinis (민물검정망둑)을 사용하는 것이 타당하다고 사료된다.

  • PDF

Development of a Simple Method to Determine the Mouse Strain from Which Cultured Cell Lines Originated

  • Yoshino, Kaori;Saijo, Kaoru;Noro, Chikako;Nakamura, Yukio
    • Interdisciplinary Bio Central
    • /
    • 제2권4호
    • /
    • pp.14.1-14.9
    • /
    • 2010
  • Misidentification of cultured cell lines results in the generation of erroneous scientific data. Hence, it is very important to identify and eliminate cell lines with a different origin from that being claimed. Various methods, such as karyotyping and isozyme analysis, can be used to detect inter-species misidentification. However, these methods have proved of little value for identifying intra-species misidentification, and it will only be through the development and application of molecular biological approaches that this will become practical. Recently, the profiling of microsatellite variants has been validated as a means of detecting gene polymorphisms and has proved to be a simple and reliable method for identifying individual cell lines. Currently, the human cell lines provided by cell banks around the world are routinely authenticated by microsatellite polymorphism profiling. Unfortunately, this practice has not been widely adopted for mouse cells lines. Here we show that the profiling of microsatellite variants can be also applied to distinguish the commonly used mouse inbred strains and to determine the strain of origin of cultured cell lines. We found that approximately 4.2% of mouse cell lines have been misidentified; this is a similar rate of misidentification as detected in human cell lines. Although this approach cannot detect intra-strain misidentification, the profiling of microsatellite variants should be routinely carried out for all mouse cell lines to eliminate inter-strain misidentification.

Molecular Characterization and Expression of LDHA and LDHB mRNA in Testes of Japanese Quail (Coturnix japonica)

  • Singh, R.P.;Sastry, K.V.H.;Pandey, N.K.;Shit, N.G.;Agarwal, R.;Singh, R.;Sharma, S.K.;Saxena, V.K.;Jagmohan, Jagmohan
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제24권8호
    • /
    • pp.1060-1068
    • /
    • 2011
  • The LDH isozymes are key catalysts in the glycolytic pathway of energy metabolism. It is well known that the distribution of the LDH isozymes vary in accordance with the metabolic requirements of different tissues. The substrates required for energy production change noticeably at successive stages of testes development suggesting a significant flexibility in the expression of glycolytic enzymes. Therefore, expression of LHDA and LDHB mRNAs was examined in adult and prepubertal quail testis. The mRNA of both LDHA and LDHB were expressed and no significant difference was observed in prepubertal testes. The mRNA levels of LDHB significantly increased during testicular development. In the adult testis, LDHA mRNA was not expressed. Expression studies revealed the presence of different LDH isozymes during testicular development. In contrast, electrophoresis of both testicular samples revealed only single band at a position indicative of an extreme type of LDH isozyme in quail testes. Furthermore, nucleotide and amino acid sequence analysis revealed significant similarity to chicken, duck and rock pigeon. These sequence results confirmed the similarity of LDHA and LDHB subunit protein in different avian species.

Constitutive Expression and Changes of Cytochrome P450 Isozymes mRNAs by Vehicles (Petrolatum, DMSO, Ethanol) in Rat Skin Using Semi-quantitative RT-PCR

  • Lee, Ai-Young;Lee, Kyung-Hoon;Ko, Duck-Sung;Chey, Won-Young
    • The Korean Journal of Physiology and Pharmacology
    • /
    • 제5권5호
    • /
    • pp.407-412
    • /
    • 2001
  • Many drugs are primarily metabolized by the cytochrome P450s (CYPs). Drug metabolites would be important allergens for adverse drug reactions such as drug eruptions. Skin tests with a suspected drug have conducted to identify causative drugs of drug eruptions, with vehicles such as white petrolatum, DMSO, ethanol. This study will compare the expression of rat CYP isozyme mRNAs between the skin and the liver, with examining an effect of the vehicles on the cutaneous CYPs using semi-quantitative RT-PCR. Thirty-two Sprague-Dawley rats between the ages of six and eight weeks were divided as four groups. One group was used to compare the constitutive mRNA expression between skin and liver, while the others were to examine the effects of three vehicles. The ratios of expression of CYP1A2, CYP2B1/2, CYP2E1, CYP3A1, and CYP4A1 were significantly higher in the liver than the skin. However, CYP1A1 and CYP2C11 were higher in the skin than liver. The effects of vehicles were quite different; white petrolatum significantly induced CYP1A1 (p=0.012) and CYP2C11 mRNAs, while ethanol inhibited CY P1A1 and CYP2B1/2. DMSO did not make any changes. The results suggest that rat skin can participate in drug metabolism with their own CYP isozymes. The effects of vehicles on the cutaneous CYP expression should not be ignored and may be applied for determination of an appropriate vehicle for certain drug(s).

  • PDF

Decreased Expression of Phospholipase C-$\beta$2 in Human Platelets with Impaired Function

  • Lee, Sang-Bong;A. Koneti Rao;Lee, Kweon-Haeng;Xu Yang;Bae, Yun-Soo
    • 한국응용약물학회:학술대회논문집
    • /
    • 한국응용약물학회 1996년도 제4회 추계심포지움
    • /
    • pp.75-84
    • /
    • 1996
  • Platelets from a patient with a mild inherited bleeding disorder and abnormal platelet aggregation and secretion show reduced generation of inositol 1,4,5-trisphosphate (IP$_3$), mobilization of intracellular Ca$\^$2+/, and phosphorylation of pleckstrin in response to several G protein mediated agonists, suggesting a possible defect at the level of phospholipase C (PLC) activation. A procedure was developed that allows quantitation of platelet PLC isozymes. After fractionation of platelet extracts by high-performance liquid chromatography, seven, out often known PLC isoforms were detected by immunoblot analysis. The amount of these isoforms in normal platelets decreased in the order PLC-${\gamma}$2 > PLC-${\beta}$2 > PLC-${\beta}$3 > PLC-${\beta}$l > PLC-${\gamma}$ > PLC-$\delta$1 > PLC-${\beta}$4. Compared with normal platelets, platelets from the patient contained approximately one-third the amount of PLC-${\beta}$2, whereas PLC-${\beta}$4 was increased threefold. These results suggest that the impaired platelet function in the patient in response to multiple G protein mediated agonists is attributable to a deficiency of PLC-${\beta}$2. They document for the first time a specific PLC isozyme deficiency in human platelets and provide an unique opportunity to understand the role of different PLC isozymes in normal platelet function.

  • PDF

까마중내 (Solanum nigrum L.) 항산화방어계의 항산화력 및 물질의 동정 (Evaluation of the Antioxidant Potential and ldentification of Active Principles of Solanum nigrum L. on Antioxidant Defense Systems)

  • 임종국;정규영;정형진
    • 생명과학회지
    • /
    • 제11권6호
    • /
    • pp.509-516
    • /
    • 2001
  • 까마중의 부위별 DPPH free radical 소거활성은 뿌리에서 가장 높았고, 줄기, 전부위, 열매 잎 그리고 꽃 순으로 높았다. 추출방법간의 DPPH free radical 소거활성은 80%methanol에 침지하여 추출한것보다는 $80^{\circ}C$에서 80% meth-anol에 흰류추출한것이 높았다. Sephades LH-20 column chromatography에 의하여 분획된 L6의 DPPH free radical 소거활성은 ethy acetate 분획 추출물보다 6.7배 높았다. 분획물내의주요물질은 2, 6-methano-3-benzazocin-11-ol, 2[1h]-pyridinethione 및 2-hydroxy-5 methylbenzaldehyde로 phenolic 화 물이었다. 줄기와 뿌리의 POD및 SOD 활성은 타 부위에 비하여 높은 활성을 나타내었고, 생육기간에는 파종 후 60일후에서 활성이 가장 높았다. PODdmlehddnl gth 패턴은 뿌리에서 10개의 band로 가장 많았고 그중 8개의 band가 식물체의 부위별로 차이가 있어Tdmaul, SOD의 동위효소 패턴은 잎에 5개의 band로 가장 많았고 그중 2개의 band가 식물체의 부위별로 차이가 있었다.이가 있었다.

  • PDF

식물육종에 있어서 비환원 ( 2n ) 배우자의 중요성 (Significance of Unreduced ( 2n ) Gametes in Plant Breeding)

  • Rim, Yong-Woo
    • 한국초지조사료학회지
    • /
    • 제17권1호
    • /
    • pp.1-10
    • /
    • 1997
  • Unreduced (2n) gametes are meiotic products (pollen or egg) having a sporophytic (somatic) chromosome number, resulting from abnormalities during either microsporogenesis or megasporogenesis. They occur naturally at a low frequency in many plant species. Unreduced (2n) gametes in plants can be identified for four possible ways as follow i) pollen size and/or shape differences between haploid (n) and diploid (2n) pollen, ii) ploidy analysis (chromosome number) of progeny or meiotic analysis (presence of dyads andlor triads at the microspore stage), iii) progeny performance and fertility and iv) dosage of isozyme and DNA markers. Unreduced (2n) gametes can be an effective breeding tool in synthesizing new cultivars, providing a unique method to maximizing heterozygosity, i.e., transferring a large proportion of the non-additive genetic effects (intra- and inter- locus interactions) h m parent to offspring and can also be used to overcome infertility of interploidy crosses. Sexual polyploidization through 2n gametes has been a major route to the formation of naturally occurring polyploids. The three mechanisms of 2n pollen formation in potato have been discovered as follow: i) parallel spindles (ps) or tripolar spindles (ts), ii) premature cytokinesis (pc-I, pc-2) and iii) synaptic mutants (sy-2, sy-3, sy-4). Genetic analysis indicated that the mechanisms of 2n gamete formation were controlled by single recessive gene in potato, alfalfa, red clover, etc., and by two recessive genes in wheat. The use of 2n gametes which can efficiently transfer germplasm fiom wild relatives to cultivated species, especially fiom diploid to tetraploid could make a contribution to the improvement of germplasm base in breeding programs.

  • PDF

Genetic Diversity Studies and Identification of Molecular and Biochemical Markers Associated with Fusarium Wilt Resistance in Cultivated Faba Bean (Vicia faba)

  • Mahmoud, Amer F.;Abd El-Fatah, Bahaa E.S.
    • The Plant Pathology Journal
    • /
    • 제36권1호
    • /
    • pp.11-28
    • /
    • 2020
  • Faba bean (Vicia faba L.) is one of the most important legume crops in Egypt. However, production of faba bean is affected by several diseases including fungal diseases. Fusarium wilt incited by Fusarium oxysporum Schlecht. was shown to be the most common wilt disease of faba bean in Assiut Governorate. Evaluation of 16 faba bean genotypes for the resistance to Fusarium wilt was carried out under greenhouse conditions. Three molecular marker systems (inter-simple sequence repeat [ISSR], sequence related amplified polymorphism [SRAP], and simple sequence repeat [SSR]) and a biochemical marker (protein profiles) were used to study the genetic diversity and detect molecular and biochemical markers associated with Fusarium wilt resistance in the tested genotypes. The results showed that certain genotypes of faba bean were resistant to Fusarium wilt, while most of the genotypes were highly susceptible. The percentage of disease severity ranged from 32.83% in Assiut-215 to 64.17% in Misr-3. The genotypes Assiut-215, Roomy-3, Marut-2, and Giza2 were the most resistant, and the genotypes Misr-3, Misr-1, Assiut-143, Giza-40, and Roomy-80 performed as highly susceptible. The genotypes Assiut-215 and Roomy-3 were considered as promising sources of the resistance to Fusarium wilt. SRAP markers showed higher polymorphism (82.53%) compared with SSR (76.85%), ISSR markers (62.24%), and protein profile (31.82%). Specific molecular and biochemical markers associated with Fusarium wilt resistance were identified. The dendrogram based on combined data of molecular and biochemical markers grouped the 16 faba bean genotypes into three clusters. Cluster I included resistant genotypes, cluster II comprised all moderate genotypes and cluster III contained highly susceptible genotypes.

${\gamma}$-Glutamyltransferase의 조직내 분포에 관한 연구 -단일클론항체의 효소면역측정법, 방사면역측정법, 면역조직화학검사, 자가방사기록검사 적용에 관하여 - (Distribution of Murine Tissue Specific ${\gamma}$-Glutamyltransferase: -Comparison of Six Monoclonal Antibody Applications in Enzyme Linked Immunosorbent Assay, Radioimmunoassay, Immunohistochemistry, and Autoradiography-)

  • 김명근;박윤규;류총근
    • 대한핵의학회지
    • /
    • 제28권1호
    • /
    • pp.112-123
    • /
    • 1994
  • ${\gamma}$-Glutamyltransferase (GGT: E.C. 2.3.2.2.) is a glycoprotein enzyme which is involved in glutathione metabolism and amino acid transport through the plasma membrane. It is distributed widely in several organs including liver and kidney. Several isozymes of GGT have been reported and some of the isozymes may be associated with hepatocarcinogenesis. We have produced six monoclnal antibodies (mAbs) against GGT purified from the liver of 2-acetamidofluorene (AAF) treated rats. All of the six mAbs were obtained by immunizing mice with liver GGT Six hybridomas which produced anti-GGT Abs were extensively subcloned and injected into the peritoneal cavity of BALB/c mice to obtain large quantities of Abs. These mAbs were purified from ascites by ammonium sulfate precipitation and protein A sepharose CL-4B column chromatography. Using these mAbs we preformed enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunohistochemistry (IHC), and autoradiography (ARG) to study the distribution of GGT isozyme in tissue. The results indicate that GGT-mAb 1 is specific for the AAF treated liver GGT, GGT-mAb 5 for the normal liver GGT, and GGT-mAb 6 for the normal kindey GGT. These mAbs may be used to evaluate the distribution of GGT isozymes in different tissues.

  • PDF

한국 약용 및 식용식물들의 항산화성 식물탐색 (Screening of Korean Medicinal and Food Plants with Antioxidant Activity)

  • 정일민;김광호;안종국
    • 한국약용작물학회지
    • /
    • 제6권4호
    • /
    • pp.311-322
    • /
    • 1998
  • 60종(種)의 자생 약용식물과 식용작물을 대상으로 항산화활성의 생리활성을 조사함으로써 약용식물과 식용작물의 유용성 측면의 확인뿐만 아니라 새로운 생리활성물질 탐색의 가능성을 검토하기 위해서 실시한 실험 결과는 다음과 같다. 자생 약용식물 및 식용작물에 대한 50% EtOH 추출물을 이용하여 TBA법. DPPH법에 의하여 1차 활성검정 결과 처리방법에 따라 활성값에 차이가 있으나 TBA법에서는 검정콩의 추출액이 87.3%, DPPH법에서는 까마중 추출액이 80.6%로서 가장 높은 활성을 보였고 두가지 방법에서 동시에 활성이 높은 것은 검정콩을 비롯하여 10종 이었다. SOD활성정도도 검정식물에 따라 큰 차이가 있으나 검정콩이 53.5%로서 가장 높은 SOD활성을 나타내었으며 SOD종류는 Cu/ZnSOD, FeSOD을 포함하는 것으로 나타났다. Cu/ZnSOD만을 함유하고 있는 잔대는 SOD활성 정도가 10.37%로서 실험 대상식물 중 가장 낮은 활성을 보였다.

  • PDF