• Title/Summary/Keyword: Marker enzyme

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A Molecular Marker Specific to Metarhizium anisopliae var. majus

  • YOON, CHEOL-SIK;GI HO SUNG;JAE MO SUNG;JAEANG OON LEE
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.334-339
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    • 1999
  • More innovative molecular markers were investigated for rapid and consistent differentiation of Metarhizium anisopliae var. majus from M. anisopliae var. anisopliae. A total of 28 isolates were obtained from various countries and hosts: 13 isolates of M. anisopliae var. anisopliae, 12 isolates of M. anisopliae var. majus, and 3 isolates of M. anisopliae collected in Korea. This study involved restriction enzyme digestions of a PCR product amplified from nuclear internally transcribed spacer (ITS) and a portion of the 28S rDNA regions. Among 11 different restriction enzymes used in this study, MboⅠ digestion particularly produced a restriction pattern that had characteristics of M. anisopliae var. majus. This restriction pattern was consistent in all isolates of M. anisopliae var. majus regardless of their geographic origins and insect hosts. Mapping experiments revealed that MboⅠ sites of M. anisopliae var. majus are identical to those of M. anisopliae var. anisopliae with an exception for the presence of an additional site in the PCR product. Results from this study provide an additional method for identification and differentiation of isolates of these two varieties of M. anisopliae for use in the field and laboratory experiments.

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Induction of Quinone Reductase and Glutathione S-Transferase in Murine Hepatoma Cells by Flavonoid Glycosides

  • Kim, Jung-Hyun;Lee, Jeong-Soon;Kim, Young-Chan;Chung, Shin-Kyo;Kwon, Chong-Suk;Kim, Young-Kyoon;Kim, Jong-Sang
    • Preventive Nutrition and Food Science
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    • v.8 no.4
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    • pp.365-371
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    • 2003
  • The potential of seven flavonoid glycosides to induce quinone reductase (QR), an anticarcinogenic marker enzyme, in murine hepatoma cells (hepalc1c7) and its mutant cells (BPRc1) was evaluated. Among test compounds, kaempferol-3-O-glucoside, luteolin-6-c-glucoside, and quercetin-3-O-glucoside (Q-3-G) induced QR in hepalc1c7 cells in a dose-dependent manner. However, in BPRc1 cells lacking arylhydrocarbon receptor nuclear translocator (ARNT), only Q-3-G caused a significant induction of quinone reductase at the concentration range of 0.5 to 8 ug/mL, suggesting that it is a monofunctional inducer. Q-3-G induced not only phase 2 enzymes, including QR and glutathione-S-transferase, but also nitroblue tetrazolium reduction activity in HL-60 cells, a biochemical marker for cell differentiation promoting agents. In conclusion, Q-3-G merits further study to evaluate its cancer chemopreventive potential.

Discrimination of Korean Agaricus bisporus cultivars using CAPS markers (CAPS 마커를 이용한 국내 개발 양송이 품종 구분)

  • Lee, Hwa-Yong;An, Hyejin;Oh, Youn-Lee;Jang, Kab-Yeul;Chung, Jong-Wook
    • Journal of Mushroom
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    • v.19 no.4
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    • pp.336-340
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    • 2021
  • The cleaved amplified polymorphic sequence (CAPS) marker uses a restriction enzyme recognition site resulting from single nucleotide polymorphisms and insertions and deletions on the DNA sequence. This technique does not require expensive equipment, the process is simple, and clear results can be obtained reliably. In this study, Agaricus bisporus cultivars SaeA, SaeDo, SaeHan, SaeYeon, SaeJeong, Dodam, Seolgang, Dahyang, Hogam, and Hadam developed in Korea were discriminated using four CAPS markers. Our results indicated that it is possible to distinguish the ten cultivars and determine the genetic diversity among them.

Characterization and Cloning of Genes Related to Embryogenic Cells in Rice - Characterization of Isozymes Related to Embryogenic Cells - (벼 배발생 세포의 특성과 배발생 관련 유전자의 분이 - 배발생 세포에 관련된 동이효소 특성 -)

  • Jung, Byung-Kyun;Paek, yun-Woong;Ko, Kyeong-Min;Nahm, Baek-Hie;Hwang, Baik
    • Journal of Plant Biology
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    • v.38 no.1
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    • pp.55-62
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    • 1995
  • Embryogenic cell (EC) suspension cultures derived from mature seed-embryo of rice (Oryza sativa L cv. Kye Hwa) were used for the expression patterns of isozyme and enzyme activity. EC suspension cultures were composed of cells that were densely cytoplasmic, potentially embryogenic. However, nonembryogenic cell (NEC) cultures were composed of large, elongated and vacuolated cells. These cells were analyzed for the isozyme pattern and enzyme activity of EC and NEC. Isozyme patterns of peroxidase, esterase, acid phosphatase and malate dehydrogenase exhibited striking difference in the total number of bands, specificity and intensity of band. Also, these isozymes showed very high activity in the EC. Specific band, band activity and higher enzyme activity of isozyme in EC was absent or low in NEC, which may indicate an association of these specific isozymes with morphological characterization and totipotency of embryogenic cells. These results indicate that specific pattern and activity of enzyme in EC could probably be used as a biochemical marker of EC in rice.n rice.

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Genetic variation of halophyte New Zealand spinach (Tetragonia tetragonioides) accessions collected in Korea using an AFLP marker (AFLP 마커를 이용한 국내수집 염생식물 번행초 유전다양성 평가)

  • Jeon, Yongsam;Jin, Yong-Tae;Choi, Seo-Hee;Park, Nuri;Kim, In-Kyung;Lee, Ka Youn;Choi, Jong-Jin;Lee, Geung-Joo
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.157-163
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    • 2016
  • This study was conducted to investigate the potential use of New Zealand spinach (Tetragonia tetragonioides) as a new vegetable crop which will be cultivated in salt-affected soils such as reclaimed areas. New Zealand spinach ecotypes native to Korea were collected across the Southern, Western and Eastern seashore regions of the Korean peninsula, among which fifty-five accessions were later further propagated and evaluated genetically by using an AFLP (amplified fragment length polymorphism) marker. Based on the AFLP analysis performed to uncover the genetic diversity of the collected ecotypes, enzymatic cleavage of the extracted DNA was implemented based on 12 EcoRI and MseI combinations. A total of 1,279 alleles (107 alleles per EcoRI and MseI enzyme combination) were successfully amplified, among which 62 alleles per enzyme combination were polymorphic (58%). The AFLP analysis indicated that the rate of genetic dissimilarity was 29% among the New Zealand spinach collections, which were clustered into the 7 genetic diversity group. This is the first report on the genetic variation in the genus Tetragonia, and the basic information can be applied to select parental lines for enhancing the segregation spectrum of the new halophytic vegetable plant grown in salt-affected areas.

분자유전학적인 기술을 이용한 육 감별법

  • Kim, Tae-Heon
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2000.11a
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    • pp.59-75
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    • 2000
  • This study was carried out to develop a DNA marker for identifying between Korean cattle (Hanwoo) and other breeds. First experiment was performed to isolate Hanwoo specific DNA marker at sequence characterized amplified regions (SCARs). Five breeds of cattle including Hanwoo, Holstein, Hereford, Angus and Charolais were represented with the from 8 to 20 individuals. Fourteen primers of 300 arbitrary primers of 10 nucleotides showed reproducible polymorphism across the breeds. An amplified band of 0.9 kb in the primer MG-3 showed the specificity to Holstein breed. And MG-6 and MG-12 detected the Hereford and Hanwoo specific markers at the size of 2.0 kb and 1.0 kb, respectively. A 1.0 kb band of MG-12 was cloned and sequenced. A SCAR primer was designed based on the obtained sequences. It was possible to identify the Hanwoo from Holstein breed. Second experiment was carried out to observe the genotype frequencies of MC1R in 1,044 samples of imported beef and eight different cattle breeds including Hanwoo, Holstein, Angus, Brown-Swiss, Charolais, Limousin, Simmental and Hereford. The primers for the amplification of bovine MC1R gene were designed based on a bovine MC1R gene sequence (GenBank accession no.Y19103). A size of 350 bp was amplified by polymerase chain reaction(PCR), digested with two different restriction enzyme, BsrFI and MspA II, and electrophoresed in 2.5% Metaphore agarose gel for determination of genotypes. Genotype frequencies of Hanwoo were 0.10 in E+e and 0.90 in ee. Allele ED was shown in all of Holstein and Angus breeds tested which have black coat color phenotypes. We suggested that SCAR marker and the bovine MC1R gene could be used as a DNA marker for distinguishing beef between Hanwoo and Holstein.

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Pepstatin- Insensitive Carboxyl Proteinase: A Biochemical Marker for Late Lysosomes in Amoeba proteus

  • Hae Kyung Kwon;HyeonJung Kim;Tae In Ahn
    • Animal cells and systems
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    • v.3 no.2
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    • pp.221-228
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    • 1999
  • In order to find a biochemical marker for late Iysosomes, we characterized two cDNAs which were cloned by using a monoclonal antibody (mAb) against Iysosomes in Amoeba proteus as a probe. The two cDNAs, a 1.3-kb cDNA in pBSK-Iys45 and a 1.6-kb cDNA in pBSK-Iys60, were found to encode proteins homologous to pepstatin-insensitive carboxyl proteinases (PICPs). E. coli transformed with pBSK-Iys45 produced two immunopositive polypeptides (45 and 43 kDa) and the cDNA in 1274 bases encoded a 44,733-Da protein (Lys45) of 420 amino acids containing one site for a core oligosaccharide. On the other hand, E. coli transformed with pBSK-Iys60 produced several polypeptides (64, 54, 45, 41, and 37 kDa) reacting with the mAb. The cDNA contained 1629 bases and encoded a 59,231-Da protein (Lys60) of 530 amino acids containing two sites for asparagine-linked core oligosaccharides. These two cDNAs showed identities of 60.3% in nucleotide sequences and 23.6% in amino acid sequences. Lys45 and Lys60 appeared to share XXEFQK as a common antigenic domain. The amino acid sequence of the Lys45 protein showed 17.4% identity and 40.9% similarity to that of PICP from Pseudomonas sp. 101. On the other hand, Lys60 showed a 24.3% identity and 51.9% similarity with human Iysosomal PICP in the amino acid sequence. A putative active center for serine protease, GTS*xxxxxFxG, was found to be conserved among PICP homologues. The two PICPs are the first reported enzymatic markers for late Iysosomes.

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Glycyrrhiza uralensis (licorice) extracts increase cell proliferation and bone marker enzyme alkaline phosphatase activity in osteoblastic MC3T3-E1 cells

  • Cho, Young-Eun;Kwun, In-Sook
    • Journal of Nutrition and Health
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    • v.51 no.4
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    • pp.316-322
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    • 2018
  • Purpose: The Glycyrrhiza uralensis species (Leguminosae) as a medicinal biocompound, and one of its root components, isoliquritigenin (ISL), which is a flavonoid, has been reported to have anti-tumor activity in vitro and in vivo. However, its function in bone formation has not been studied yet. In this study, we tested the effect of Glycyrrhiza uralensis (ErLR) and baked Glycyrrhiza uralensis (EdLR) extracts on osteoblast proliferation, alkaline phosphatase (ALP) activity, and bone-related gene expression in osteoblastic MC3T3-E1 cells. Methods: MC3T3-E1 cells were cultured in various levels of ErLR (0, 5, 10, 15, $20{\mu}g/mL$), EdLR (0, 5, 10, 15, $20{\mu}g/mL$), or ISL (0, 5, 10, 15, $20{\mu}M$) in time sequences (1, 5, and 20 days). Also, isoliquritigenin (ISL) was tested for comparison to those two biocompound extracts. Results: MTT assay results showed that all three compounds (ErLR, EdLR, and ISL) increased osteoblastic-cell proliferation in a concentration-dependent manner for one day. In addition, both ErLR and EdLR compounds elevated the osteoblast proliferation for 5 or 20 days. Extracellular ALP activity was also increased as ErLR, EdLR, and ISL concentration increased at 20 days, which implies the positive effect of Glycyrrhiza species on osteoblast mineralization. The bone-related marker mRNAs were upregulated in the ErLR-treated osteoblastic MC3T3-E1 cells for 20 days. Bone-specific transcription factor Runx2 gene expression was also elevated in the ErLR- and EdLR-treated osteoblastic MC3T3-E1 cells for 20 days. Conclusion: These results demonstrated that Glycyrrhiza uralensis extracts may be useful for preventing osteoporosis by increasing cell proliferation, ALP activity, and bone-marker gene expression in osteoblastic cells.

Development of CAPS marker for identifying a Formosan lily (Lilium formosanum) (흰나리(Lilium formosanum Wallace) 식별을 위한 CAPS 마커의 개발)

  • Chung, Sung Jin;Lee, Ka Youn;Yoon, A Ra;Jang, Ji Young;Kim, Jin Kug;Lee, Geung-Joo
    • Korean Journal of Agricultural Science
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    • v.41 no.2
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    • pp.101-106
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    • 2014
  • This study was conducted to identify lily species native to Korea from formosan lily (Lilium formosanum) belonging to Longiflorum section. Due to flowering time, flower color and orientation, long shelf life and resistant to diseases, the native lily species can be valuable genetic resources for interspecific hybrids. One of the chloroplast genes, matK, was used to clone and sequence to explore any base changes. The matK was successfully amplified into 1,539 bp (94% of the gene) and phylogenetic tree demonstrated 6 clades for those 11 lily species used in this study. There were one or two base substitutions among 10 lilies native to Korea, while formosan lily native to Taiwan exhibited 6 base substitutions in matK gene, rendering it genetically distant. A restriction enzyme NruI recognized one of the six base changes, and digested the matK gene of 10 native lily species only, but not in formosan lily. The confirmed cleavage characteristic of the target region in matK gene was designed into a CAPS (cleaved amplified polymorphic sequences) marker which will be available to estimate compatibility of interspecific hybridization and to trace the pedigree when those native lilies are crossed with the formosan lily.

Relationship between Preoperative Serum CA15-3 and CEA Levels and Clinicopathological Parameters in Breast Cancer

  • Moazzezy, Neda;Farahany, Tahereh Zarnoosheh;Oloomi, Mana;Bouzari, Saeid
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1685-1688
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    • 2014
  • Background: CEA and CA 15.3 serum tumor markers are currently used in clinical practice for monitoring therapy. The aim of this study was to evaluate serum level of these markers among healthy females and invasive breast carcinoma (IBC) patients and to determine any relationships with clinicopathological factors. Materials and Methods: 60 Iranian females were enrolled in this study, 30 healthy and 30 diagnosed with breast cancer who had not received any preoperative chemotherapy or hormone therapy. Enzyme linked immunosorbent assays were used for the quantitative determination of the cancer associated antigens, CEA and MUC1 (CA15-3). Results: The serological levels of CEA and CA15-3 ($5.0033{\pm}0.49{\mu}g/L$ and $178.1667{\pm}15.11$ U/ml) in the breast cancer patients were significantly higher (p=0.00) than the serum levels of normal controls ($1.1237{\pm}0.11{\mu}g/L$ and $21.13{\pm}3.058$ U/ml). Regarding the CEA marker, a significant correlation with grade of tumor was shown. Furthermore, there was a low correlation between CA15-3 and CEA marker with correlation coefficient r=0.08. Conclusions: Collectively, markedly high levels of CEA and CA15-3 were found in our patients, pointing to their use as additional tools after clinical diagnosis.