• 제목/요약/키워드: DNA variants

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Effect of green tea extract in extender of Simmental bull semen on pregnancy rate of recipients

  • Susilowati, Suherni;Sardjito, Trilas;Mustofa, Imam;Widodo, Oky Setio;Kurnijasanti, Rochmah
    • Animal Bioscience
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    • 제34권2호
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    • pp.198-204
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    • 2021
  • Objective: The aim of this study was to ascertain the effects of adding green tea extract (GTE) to skim milk-egg yolk (SM-EY) extender on both the quality of post-thawed bull semen and the pregnancy rates of the recipient cows. Methods: Twelve ejaculates from four Simmental bulls, aged 3 to 5 years and weighing 900 to 950 kg, were diluted SM-EY extender, added with 0, 0.05, 0.1, and 0.15 mg GTE/100 mL extender and then frozen. After four weeks storage in liquid nitrogen, the sperm were thawed and evaluated for viability, motility, intact plasma membrane (IPM), and DNA fragmentation. Meanwhile, the estrus cycles of 48 recipient cows were synchronized by intramuscular administration of a single injection of 5 mg prostaglandin F2α. Estrus cows were divided into four equal groups and inseminated artificially 18 to 20 h after the onset of estrus by using semen from each extender group. Pregnancy was diagnosed by measuring serum progesterone levels at 21 days, followed by transrectal palpation 90 days after insemination. Results: The findings revealed that adding 0.1 mg of GTE/100 mL extender produced the highest percentages of sperm viability (70.67%±1.75%), motility (69.17%±1.47%), and IPM (69.23%±1.21%) and the lowest percentage of DNA fragmentation (3.00%±0.50%). The pregnancy diagnosis revealed that all cows (36/36) inseminated using frozen semen in GTE addition extender were pregnant (pregnancy rate 100%), whereas the pregnancy rate of the control group was 83.33% (10/12). Conclusion: It may be concluded that 0.1 mg GTE/100 mL extender yields the best quality of spermatozoa and that all variants doses of GTE in extender produce a higher pregnancy rate among recipient cows.

한국인의 피부 기저세포암종과 편평세포암종의 XRCC1 유전자 다형 (Polymorphism in the DNA Repair Gene XRCC1 Associated with Squamous Cell Carcinoma and Basal Cell Carcinoma of the Skin in Koreans)

  • 강상윤;이광길;심정연;정윤규;김남근;민완기
    • Archives of Plastic Surgery
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    • 제33권4호
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    • pp.433-439
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    • 2006
  • Purpose: DNA in most cell is regularly damaged by endogenous and exogenous mutagens. Unrepaired damage resulted in apoptosis or may lead to unregulated cell growth and cancer. Inheritance of genetic variants at one or more loci results in an reduced DNA repair capacity. These polymorphisms are highly prevalent in the population, and therefore the attributable risks for cancer could be high. Several studies have documented that polymorphisms of XRCC1, XPD and XRCC3 are associated with skin cancer, especially, XRCC1 among of them has been reported frequently. So, this study involves the relationship between mutation of XRCC1 of squamous cell and basal cell cancer of the skin and risk of cancer development in Korean population. Methods: In case control study, study population (n=100, each cancer) is patients who were pathologically diagnosed as skin cancer(squamous cell carcinoma and basal cell carcinoma) in Yonsei Wonju Christian Hospital and Bundang CHA General Hospital between 1998 and 2004. The samples of DNA from whom no history of premalignant skin lesion and other malignant diseases were reported belonged to the control group(n=210). Blood and tissue samples were analyzed for presence of XRCC1 Arg399Glu, Arg280His, Arg194Trp using PCR/ RFLP method. Results: For Korean, there was a significant correlation between XRCC1 Arg399Gln gene mutation and risk of basal cell carcinoma development(Arg 399Gln(GA), p=0.012, OR=2.016, 95% CI; 1.230-3.305) /Arg399Gln (AA), p=0.011, OR=1.864, 95% CI; 1.149-3.026)). And, there was also significant correlation between XRCC1 Arg194Trp and risk of skin squamous cell carcinoma development (Arg194Trp (CT+TT), p=0.041, OR=0.537, 95% CI; 0.301-0.960)). In contrast, there was no significant correlation between XRCC1 Arg280His and risk of either basal cell carcinoma or squamous cell carcinoma development. Conclusions: Our result present that XRCC1 Arg399 Gln in basal cell carcinoma and XRCC1 Arg194Trp in squamous cell carcinoma have possibility of cancer risk and biomarker in Korean population. But XRCC1 Arg280 His known having cancer risk on other studies is not associated with cancer risk to squamous cell carcinoma and basal cell carcinoma in Korean population.

Studies of Opsin Genes in a Smelt Fish, Ayu (Plecoglossus altivelis)

  • Minamoto, Toshifumi;Shimizu, Isamu
    • Journal of Photoscience
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    • 제9권2호
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    • pp.269-271
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    • 2002
  • To investigate the visual and extra-ocular photoreception, we cloned the opsin genes in ayu (Plecoglossus allivelis). Amplified fragments encoding exon-4 (-5) of opsin cDNAs were cloned from the retina and brains of ayu, and sequenced. One clone was identified as rod (AYU-Rh), two as green cone (AYU-GI, -G2), one as red cone (A YU-R), two as ultraviolet cone (AYU-UVl, UV2), one as VA (AYU-VA), and one as extra-ocular rod (AYU-ExoRh) opsins. 335 amino acids sequence deduced from the full-length cDNA of AYU-Rh showed high identity with that of other fish. Southern blotting analysis indicated that ayu possess two 'rhodopsin' genes, one is visual rhodopsin and the other is non-visual extra-ocular rhodopsin. In situ hybridization showed that the mRNA of AYU-Rh was localized only in rod cells in the retina. On the other hands, AYU-ExoRh was expressed only in the pineal. We cloned two isoforms (AYU-VAM and -VAL) of VA opsin from ayu. The deduced amino acid sequences of these variants were identical to each other within the first 342 residues, but they showed divergence in the C-terminal sequence. AYU- VAL corresponded to the long isoform found in other fish, and AYU-VAM was identified as a new type of VA opsin variant. Pal-VAM is a new probably functional non-visual photoreceptive molecule in fish.

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Opportunities and Challenges in Nutrigenomics and Health Promotion

  • Milner John A.
    • 한국식품영양과학회:학술대회논문집
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    • 한국식품영양과학회 2004년도 Annual Meeting and International Symposium
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    • pp.17-23
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    • 2004
  • Not all individuals respond identically, or at times in the same direction, to dietary interventions. These inconsistencies likely arise because of diet and genomic interactions (nutrigenomics effects). A host of factors may influence the response to bioactive food components including specific polymorphisms (nutrigenetic effect), DNA methylation patterns and other epigenomic factors (nutritional epigenomic effects), capacity to induce anuo. suppress specific mRNA expression and patterns (nutritional transcriptomics), the occurrence and activity of proteins (proteomic effects), and/or the dose and temporal changes in cellular small molecular weight compounds will not only provide clues about specificity in response to food components, but assist in the identification of surrogate tissues and biomarkers that can predict a response. While this 'discovery' phase is critical for defining mechanisms and targets, and thus those who will benefit most from intervention, its true usefulness depends on moving this understanding into 'development' (interventions for better prevention, detection, diagnosis, and treatment) and a 'delivery' phase where information is provided to those most in need. It is incumbent on those involved with food and nutrition to embrace the 'omics' that relate to nutrition when considering not only the nutritional value of foods and their food components, but also when addressing acceptability and safety. The future of 'Nutrigenomics and Health Promotion' depends on the ability of the scientific community to identity appropriate biomarkers and susceptibility variants, effective communications about the merits of such undertakings with the health care community and with consumers, and doing all of this within a responsible bioethical framework.

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Functional Characterization of 5-Enopyruvylshikimate-3-Phosphate Synthase from Alkaliphilus metalliredigens in Transgenic Arabidopsis

  • Xing, Xiao-Juan;Tian, Yong-Sheng;Peng, Ri-He;Xu, Jing;Zhao, Wei;Yao, Quan-Hong;Sun, Sheng
    • Journal of Microbiology and Biotechnology
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    • 제24권10호
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    • pp.1421-1426
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    • 2014
  • Although a large number of AroA enzymes (EPSPS: 5-enopyruvylshikimate-3-phosphate synthase) have been identified, cloned, and tested for glyphosate resistance, only two AroA variants, derived from Agrobacterium tumefaciens strain CP4 and Zea mays, have been utilized to produce the commercial glyphosate-resistant crops. Here, we have used a PCR-based twostep DNA synthesis method to synthesize an aroA gene ($aroA_{A.\;metalliredigens}$) from Alkaliphilus metalliredigens, encoding a new EPSPS. Furthermore, transgenic Arabidopsis with the new $aroA_{A.\;metalliredigens}$ gene was obtained to confirm the potential of the novel aroA gene in developing glyphosate-resistant crops.

Putative Histone H2A Genes from a Red Alga, Griffithsia japonica

  • Lee, Yoo-Kyung;Lee, Hong-Kum
    • ALGAE
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    • 제18권3호
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    • pp.191-197
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    • 2003
  • Histones are important proteins that interact with the DNA double helix to form nucleosome. Two putative histone genes, GjH2A-1 and GjH2A-2 were isolated from a red alga Griffithsia japonica. The putative open reading frame of GjH2A-1 and GjH2A-2 shared high similarity with the previously reported amino acid sequences of histone H2As. They have a motif consisting of seven amino acids A-G-L-Q-F-P-V, which matches the histone H2A motif [AC]-G-L-x-F-P-V. Phylogenetic trees were constructed from amino acid sequences of 38 histone H2As. The histone H2As were divided into two groups: major H2As and H2A.F/Z variants. The major histone H2A group consisted of animals, fungi, plants + green algae, and red algae H2A subgroups. The animal histone H2A subgroup was divided into vertebrates, echinoderms, nematodes, insects, and segmented worms H2As. The putative red algal histone genes, GjH2A-1 and GjH2A-2, constituted an independent lineage. This is the first report on red algal histone genes.

A Neutravidin-based Assay for Reverse Transcriptase Suitable for High Throughput Screening of Retroviral Activity

  • Brennan, Lyndall E.;Sune, Carlos;Klimkait, Thomas
    • BMB Reports
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    • 제35권3호
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    • pp.262-266
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    • 2002
  • A non-isotopic neutravidin-based reverse transcriptase (RT) assay adapted for high throughput screening of HIV activity is described. Using a 96-well microtitre plate, HIV particles are lysed and the RT enzyme released into a reaction mixture containing poly(A) RNA, biotinylated oligo d(T) and fluorescein-labelled dUTP (FI-dUTP). With poly(A) as a template and oligo d(T) as primer, the viron RT incorporates FI-dUTP into an elongating DNA strand. The resulting product is captured on a neutravidin-coated 96-well plate and the unincorporated nucleotides removed by a series of washing steps. A simple ELISA is subsequently performed using a monoclonal antifluorescein antibody conjugated to alkaline phosphatase. Quantification of RT activity is facilitated by a colorimetric readout. The assay was validated in the context of a diagnostic HIV-1 phenotyping assay. Using supernatants from HIV-1 infected lymphocyte cultures the assay was shown to be as sensitive as a radioactive assay and the RT activity correlated well with levels of cell-asociated HIV-p24. Importantly, even minor reductions of RT activity by virus variants with reduced fitness could be distinguished.

A Review of Detection Methods for the Plant Viruses

  • Jeong, Joo-Jin;Ju, Ho-Jong;Noh, Jaejong
    • 식물병연구
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    • 제20권3호
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    • pp.173-181
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    • 2014
  • The early and accurate detection of plant viruses is an essential component to control those. Because the globalization of trade by free trade agreement (FTA) and the rapid climate change promote the country-to-country transfer of viruses and their hosts and vectors, diagnosis of viral diseases is getting more important. Because symptoms of viral diseases are not distinct with great variety and are confused with those of abiotic stresses, symptomatic diagnosis may not be appropriate. From the last three decades, enzyme-linked immunosorbent assays (ELISAs), developed based on serological principle, have been widely used. However, ELISAs to detect plant viruses decrease due to some limitations such as availability of antibody for target virus, cost to produce antibody, requirement of large volume of sample, and time to complete ELISAs. Many advanced techniques allow overcoming demerits of ELISAs. Since the polymerase chain reaction (PCR) developed as a technique to amplify target DNA, PCR evolved to many variants with greater sensitivity than ELISAs. Many systems of plant virus detection are reviewed here, which includes immunological-based detection system, PCR techniques, and hybridization-based methods such as microarray. Some of techniques have been used in practical, while some are still under developing to get the level of confidence for actual use.

In Vitro Regeneration of Lycium chinense Miller and Detection of Silent Somaclones with RAPD Polymorphisms

  • Ahn, In-Suk;Park, Young-Goo;Shin, Dong-Ill;Sul, Ill-Whan
    • Journal of Plant Biotechnology
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    • 제6권3호
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    • pp.157-163
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    • 2004
  • An efficient system for the regeneration of adventitious shoots from in vitro cultured leaf sections of Lycium chinense Miller was developed and silent somaclones from the regenerants detected with RAPD method. Among the eight media tested (B5, SH, N&N, 1/2MS, MS, 3/2MS, GD and WPM), and four cytokinins (BA, kinetin, 2ip and zeatin) with different concentrations (1, 5, 10, 20, 30 and 40 $\mu{M}$), 1/2 MS medium supplemented with 20 and 30 $\mu{M}$ zeatin showed the best regeneration frequency (100% and 93.7%) and higher average number of shoots (9.0 and 9.4). All regenerants easily elongated after subculturing on 1/4MS without growth stimulants and produced spontaneous adventitious roots from their basal parts. With phenotypically normal 40 regenerants, RAPD analysis with 15 different random primers was performed to examine the cryptic somaclonal variants. No substantial differences in banding patterns were found in the amplified polymorphic DNAs implying no DNA changes during dedifferentiation into adventitious shoots. However, one (OPF-4) of the 15 primers detected silent somaclonal variation in one regenerant in which two different polymorphic bands did not appear when compared with the rest regenerants. The results indicate that regenerantion via intervening callus phase can be used to establish true-to-type planting stocks for homogeneous population.

Generation of Whole-Genome Sequencing Data for Comparing Primary and Castration-Resistant Prostate Cancer

  • Park, Jong-Lyul;Kim, Seon-Kyu;Kim, Jeong-Hwan;Yun, Seok Joong;Kim, Wun-Jae;Kim, Won Tae;Jeong, Pildu;Kang, Ho Won;Kim, Seon-Young
    • Genomics & Informatics
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    • 제16권3호
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    • pp.71-74
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    • 2018
  • Because castration-resistant prostate cancer (CRPC) does not respond to androgen deprivation therapy and has a very poor prognosis, it is critical to identify a prognostic indicator for predicting high-risk patients who will develop CRPC. Here, we report a dataset of whole genomes from four pairs of primary prostate cancer (PC) and CRPC samples. The analysis of the paired PC and CRPC samples in the whole-genome data showed that the average number of somatic mutations per patients was 7,927 in CRPC tissues compared with primary PC tissues (range, 1,691 to 21,705). Our whole-genome sequencing data of primary PC and CRPC may be useful for understanding the genomic changes and molecular mechanisms that occur during the progression from PC to CRPC.