• Title/Summary/Keyword: 서열화

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Construction and In vitro Study of a Prx 6/Luc Vector System for Screening Antioxidant Compounds in the Transgenic Mice (항산화반응을 유발하는 물질의 검색에 적용할 수 있는 형질전환 마우스 생산을 위한 새로운 Prx 6/Luc 벡터시스템의 제조 및 폐암세포주에서 반응성 확인)

  • Lee, Young Ju;Nam, So Hee;Kim, Ji Eun;Hwang, In Sik;Lee, Hye Ryun;Choi, Sun Il;Kwak, Moon Hwa;Lee, Jae Ho;Jung, Young Jin;An, Beum Soo;Hwang, Dae Youn
    • Journal of Life Science
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    • v.23 no.2
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    • pp.167-174
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    • 2013
  • Peroxiredoxin 6 (Prx 6) is a member of the thiol-specific antioxidant protein family, which may play a role in protection against oxidative stress and in regulating phospholipid turnover. The aim of this study was to determine whether a human Prx 6/Luc vector was stably expressed and responded to antioxidants in a lung cell line (NCI-H460). To achieve this, the luciferase signal, hPrx 6 mRNA expression, and superoxide dismutase (SOD) activity were measured in transfectants with a hPrx 6/Luc plasmid after treatment with four antioxidant extracts, including Korea white ginseng (KWG), Korea red ginseng (KRG), Liriope platyphylla (LP), and red Liriope platyphylla (RLP). First, the hPrx 6/Luc plasmid was successfully constructed with DNA fragments of human Prx 6 promoter, amplified by PCR using genomic DNA isolated from NCI-H460 cells, and cloned into the pTransLucent reporter vector. The orientation and sequencing of the hPrx 6/Luc plasmid were identified with restriction enzyme and automatic sequencing. A luciferase assay revealed significant enhancement of luciferase activity in the four treatment groups compared with a vehicle-treated group, although the ratio of the increase was different within each group. The KRG- and LP-treated groups showed higher activity than the KWG- and RLP-treated groups. Furthermore, the luciferase activity against RLP occurred roughly in a dose-dependent manner. However, the level of endogenous hPrx 6 mRNA did not change in any group treated with the four extracts. The SOD activity was in agreement with the luciferase activity. Therefore, these results indicate that the hPrx 6/Luc vector system may successfully express and respond to antioxidant compounds in NCI-H460 cells. The data also suggest that the Prx 6/Luc vector system may be effectively applied in screening the response of hPrx 6 to antioxidant compounds in transgenic mice.

Analysis of Mutant Chinese Cabbage Plants Using Gene Tagging System (Gene Tagging System을 이용한 돌연변이 배추의 분석)

  • Yu, Jae-Gyeong;Lee, Gi-Ho;Lim, Ki-Byung;Hwang, Yoon-Jung;Woo, Eun-Taek;Kim, Jung-Sun;Park, Beom-Seok;Lee, Youn-Hyung;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.28 no.3
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    • pp.442-448
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    • 2010
  • The objectives of this study were to analyze mutant lines of Chinese cabbage ($Brassica$ $rapa$ ssp. $pekinensis$) using gene tagging system (plasmid rescue and inverse polymerase chain reaction) and to observe the phenotypic characteristics. Insertional mutants were derived by transferring DNA (T-DNA) of $Agrobacterium$ for functional genomics study in Chinese cabbage. The hypocotyls of Chinese cabbage 'Seoul' were used to obtain transgenic plants with $Agrobacterium$ $tumefaciens$ harboring pRCV2 vector. To tag T-DNA from the Chinese cabbage genomic DNA, plasmid rescue and inverse PCR were applied for multiple copies and single copy insertional mutants. These techniques were successfully conducted to Chinese cabbage plant with high efficiency, and as a result, T-DNA of pRCV2 vector showed distinct various integration patterns in the transgenic plant genome. The polyploidy level analysis showed the change in phenotypic characteristics of 13 mutant lines was not due to variation in somatic chromosome number. Compared with wild type, the $T_1$ progenies showed varied phenotypes, such as decreased stamen numbers, larger or smaller flowers, upright growth habit, hairless leaves, chlorosis symptoms, narrow leaves, and deeply serrated leaves. The polyploidy level analysis showed the change in phenotypic characteristics of 13 mutant lines was not due to variation in somatic chromosome number. To tag T-DNA from the Chinese cabbage genomic DNA, plasmid rescue and inverse PCR were applied for multiple copies and single copy insertional mutants. Mutants that showed distinct phenotypic difference compared to wild type with 1 copy of T-DNA by Southern blot analysis, and with 2n = 20 of chromosome number were selected. These selected mutant lines were sequenced flanking DNA, mapped genomic loci, and the genome information of the lines is being recorded in specially developed database.

Identification of Compound Heterozygous Alleles in a Patient with Autosomal Recessive Limb-Girdle Muscular Dystrophy (상염색체 열성 지대형 근이영양증 환자로부터 TTN 유전자의 복합 이형접합성 대립유전자의 분리)

  • Choi, Hee Ji;Lee, Soo Bin;Kwon, Hye Mi;Choi, Byung-Ok;Chung, Ki Wha
    • Journal of Life Science
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    • v.31 no.10
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    • pp.913-921
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    • 2021
  • Limb-girdle muscular dystrophy (LGMD) which is characterized by progressive muscle weakening of the hip and shoulder shows both dominant and recessive inheritances with many pathogenic genes including TTN. This study performed to identify genetic causes of a male patient with late onset (45 years old) autosomal recessive LGMD and atrial flutter. By application of the whole exome sequencing, we identified bi-allelic variants of TTN gene in the patient. One allele had a single missense variant of [c.24124G>T (p.V8042F)], while the other allele consisted of three missense variants of [c.29222G>C (p.R9741P) + c.67490A>G (p.H22497R) + c.75376C>T (p.R25126C)]. The p.V8042F allele was transmitted from his mother, while the other haplotype allele was putatively transmitted from his father. His two unaffected sons had only the p.R9741P. These variants have been not reported or rarely reported in the public human genome databases (1,000 Genome, gnomAD, and KRGDB). Most variants were located in the highly conserved immunoglobulin or fibronectin domains and were predicted to be pathogenic by the in silico analyses. The TTN giant protein plays a key role in muscle assembly, force transmission at the Z-line, and maintenance of resting tension in the I-band. In conclusion, we think that these bi-allelic compound heterozygous mutations may play a role as the genetic causes of the LGMD phenotype.

An essay on appraisal method over official administration records ill-balanced. -For development of appraisal process and method over chosun government-general office records- (불균형 잔존 행정기록의 평가방법 시론 - 조선총독부 공문서의 평가절차론 수립을 위하여 -)

  • Kim, Ik-Han
    • The Korean Journal of Archival Studies
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    • no.13
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    • pp.179-203
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    • 2006
  • This study develops the process and method of official administration documents which have remained ill-balanced like the official documents of the government-general of Chosun(the pro-Japanese colonial government (1910-1945)). At first, the existing Appraisal-theories are recomposed. The Appraisal-Theories of Schellenberg is focused valuation about value of records itself, but fuction-Appraisal theory is attached importance to operational activities which take the record into action. But given that the record is a re-presentation of operational activities, the both are the same on the philosophy aspect. Therefore, in the case that the process - method is properly designed, it can be possible to use a composite type between operational activities and records. Also, a method of the Curve has its strong points in the macro and balanced aspect while the Absolute has it's strength in the micro aspect, so that chances are that both alternate methodologies are applied to the study. Hereby, the existing Appraisal theories are concluded to be the mutually-complemented things that can be easily put together into various forms according to the characteristics of an object and its situation, in the terms of the specific Appraisal methodology. Especially, in the case of this article dealing with the imbalance remains official-documents, it is necessary to compromise more properly process with a indicated useful method than establishing a method and process by choosing the only one theory. In order to appraise the official-documents of the pro-Japanese colonial government (1910-1945), a macro appraisal of value has to be appraised about them by understanding a system, functions and using the historical-cultural evolution, after analysing Disposal Authority. From this, map the record so that organization function maps are constructed regarding the value rank of functions and detailed-functions. After this, establish the appraisal strategy considering the internal environment of archival agencies and based on micro appraisal to a great quantity of records remained and supplying other meaning to a small quantity of records remained for example, the oral resources production are accomplished. The study has not yet reached the following aspects ; a function analysis, historical decoding techniques, a curve valuation of the record, the official gazette of the government general of Chosun( the pro-Japanese government for 1910-1945), an analysis method of the other historical materials and it's process, presentation of appraisal output image. As the result, that's just simply a proposal and we should fill in the above-mentioned shortages of the study through development of all the up-coming studies.

Complete genome sequence of Lachnospiraceae bacterium KGMB03038 (=KCTC 15821) isolated from healthy Korean feces (건강한 한국인 분변으로부터 분리된 Lachnospiraceae bacterium KGMB03038 (=KCTC 15821) 균주의 유전체 염기서열 초안)

  • Kim, Ji-Sun;Kang, Se Won;Han, Kook-Il;Lee, Keun Chul;Eom, Mi Kyung;Suh, Min Kuk;Kim, Han Sol;Lee, Ju Huck;Park, Seung-Hwan;Park, Jam-Eon;Oh, Byeong Seob;Yu, Seung Yeob;Choi, Seung-Hyeon;Ryu, Seoung Woo;Lee, Dong Ho;Yoon, Hyuk;Kim, Byung-Yong;Lee, Je Hee;Lee, Jung-Sook
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.289-292
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    • 2019
  • Lachnospiraceae bacterium KGMB03038 (=KCTC 15821) belonging to the class Clostridia in phylum Firmicutes, was isolated from a stool sample of a healthy Korean. Herein, we report the complete genome sequence of strain KGMB03038 analyzed using the PacBio Sequel platform. The genome comprises of 3,334,474 bp with G + C content of 47.8%, which includes 3,099 predicted protein-coding genes, 12 ribosomal RNAs, 54 transfer RNAs, and 4 ncRNAs. Genome analysis revealed that strain KGMB03038 possesses a number of genes involved in hydrolysis of carbohydrates, including mono-, di-, and oligo-saccharides, and biosynthesis of various amino acids.

Identification of Homozygous Mutations in Two Consanguineous Families with Hearing Loss (청력 장애를 나타내는 두 근친 가계로부터 동형접합성 돌연변이의 분리)

  • Lim, Si On;Park, Hye Ri;Jung, Na Young;Park, Cho Eun;Kanwal, Sumaira;Chung, Ki Wha
    • Journal of Life Science
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    • v.31 no.5
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    • pp.453-463
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    • 2021
  • Hearing loss is a group of clinically and genetically heterogeneous disorders characterized by congenital- to adult-onset deafness with frequent additional symptoms such as myopathy, nephropathy, and optic disorders. It is commonly divided into two types: syndromic, with no other symptoms, and nonsyndromic, with other symptoms. Autosomal recessive hearing loss is relatively frequent in Pakistan, which may be due in part to frequent consanguineous marriages. This study was performed by whole exome sequencing to determine the genetic causes in two Pakistani consanguineous families with autosomal recessive hearing loss. We identified a pathogenic homozygous variant (p.Leu326Gln in MYO7A) in a family with prelingual-onset hearing loss and two variants of uncertain significance (p.Val3094Ile in GPR98 and p.Asp56Gly in PLA2G6) in a family with early-onset hearing loss concurrent with muscular atrophy. The missense mutations in MYO7A and PLA2G6 were located in the highly conserved sites, and in silico analyses predicted pathogenicity, while the GPR98 mutation was located in the less conserved site, and most in silico analysis programs predicted its nonpathogenic effect. Homozygosity mapping showed that both alleles of the homozygous mutations identified in each family originated from a single founder; spread from this single source might be due to consanguineous marriages. This study will help provide exact molecular diagnosis and treatment for autosomal recessive hearing loss patients in Pakistan.

Development of Method using LC-ESI-MS/MS and KASP for Identification of Gymnema sylvestre in Food (식품에서 당살초 판별을 위한 LC-ESI-MS/MS 분석법과 KASP 마커 개발)

  • Park, Boreum;Lee, Sun Hee;Eom, Kwonyong;Noh, Eunyoung;Moon Han, Kyoung;Hwang, Jinwoo;Kim, Hyungil;Baek, Sun Young
    • Journal of Food Hygiene and Safety
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    • v.37 no.2
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    • pp.46-54
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    • 2022
  • Known for its effectiveness in weight loss and diabetes prevention, Gymnema sylvestre products can be found in the US, Japanese, and Indian markets. However, the recommended dosage or safety of these products has not yet been proven. Therefore, development of an analytical method for detecting the content of Gymnema sylvestre in food products is required. Accordingly, this study proposes an analysis method that can examine Gymnema sylvestre in food using LC-ESI-MS/MS and KASP (Kompetitive Allele-Specific PCR) markers. In LC-ESI-MS/MS, a simultaneous analysis method for gymnemic acid and deacylgymnemic acid was optimized using negative ionization mode, and its validation test was completed for solid and liquid samples. In addition, KASP markers were prepared by finding the specific SNP of G. sylvestre in ITS2 and matK through DNA barcodes. The two KASP markers returned positive FAM fluorescence result when combined with G. sylvestre, and this aspect was confirmed in raw G. sylvestre as well. The applicability of the method was tested on 21 different food and healthy functional products containing G. sylvestre purchased on the internet. As a result, although there was a difference in the ratios of gymnemic acid and deacylgymnemic acid in LC-ESI-MS/MS, the index component was detected in all 21 products samples. In the KASP analysis, 9 products returned positive FAM result, and the rest of the products were found to be containing G. sylvestre extract. This study is the first study to use the dual system of LC-ESI-MS/MS and KASP for the analysis of G. sylvestre. The study has confirmed that these two methods are applicable to the examine G. sylvestre content in food products.

Review of the Korean Indigenous Species Investigation Project (2006-2020) by the National Institute of Biological Resources under the Ministry of Environment, Republic of Korea (한반도 자생생물 조사·발굴 연구사업 고찰(2006~2020))

  • Bae, Yeon Jae;Cho, Kijong;Min, Gi-Sik;Kim, Byung-Jik;Hyun, Jin-Oh;Lee, Jin Hwan;Lee, Hyang Burm;Yoon, Jung-Hoon;Hwang, Jeong Mi;Yum, Jin Hwa
    • Korean Journal of Environmental Biology
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    • v.39 no.1
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    • pp.119-135
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    • 2021
  • Korea has stepped up efforts to investigate and catalog its flora and fauna to conserve the biodiversity of the Korean Peninsula and secure biological resources since the ratification of the Convention on Biological Diversity (CBD) in 1992 and the Nagoya Protocol on Access to Genetic Resources and the Fair and Equitable Sharing of Benefits (ABS) in 2010. Thus, after its establishment in 2007, the National Institute of Biological Resources (NIBR) of the Ministry of Environment of Korea initiated a project called the Korean Indigenous Species Investigation Project to investigate indigenous species on the Korean Peninsula. For 15 years since its beginning in 2006, this project has been carried out in five phases, Phase 1 from 2006-2008, Phase 2 from 2009-2011, Phase 3 from 2012-2014, Phase 4 from 2015-2017, and Phase 5 from 2018-2020. Before this project, in 2006, the number of indigenous species surveyed was 29,916. The figure was cumulatively aggregated at the end of each phase as 33,253 species for Phase 1 (2008), 38,011 species for Phase 2 (2011), 42,756 species for Phase 3 (2014), 49,027 species for Phase 4 (2017), and 54,428 species for Phase 5(2020). The number of indigenous species surveyed grew rapidly, showing an approximately 1.8-fold increase as the project progressed. These statistics showed an annual average of 2,320 newly recorded species during the project period. Among the recorded species, a total of 5,242 new species were reported in scientific publications, a great scientific achievement. During this project period, newly recorded species on the Korean Peninsula were identified using the recent taxonomic classifications as follows: 4,440 insect species (including 988 new species), 4,333 invertebrate species except for insects (including 1,492 new species), 98 vertebrate species (fish) (including nine new species), 309 plant species (including 176 vascular plant species, 133 bryophyte species, and 39 new species), 1,916 algae species (including 178 new species), 1,716 fungi and lichen species(including 309 new species), and 4,812 prokaryotic species (including 2,226 new species). The number of collected biological specimens in each phase was aggregated as follows: 247,226 for Phase 1 (2008), 207,827 for Phase 2 (2011), 287,133 for Phase 3 (2014), 244,920 for Phase 4(2017), and 144,333 for Phase 5(2020). A total of 1,131,439 specimens were obtained with an annual average of 75,429. More specifically, 281,054 insect specimens, 194,667 invertebrate specimens (except for insects), 40,100 fish specimens, 378,251 plant specimens, 140,490 algae specimens, 61,695 fungi specimens, and 35,182 prokaryotic specimens were collected. The cumulative number of researchers, which were nearly all professional taxonomists and graduate students majoring in taxonomy across the country, involved in this project was around 5,000, with an annual average of 395. The number of researchers/assistant researchers or mainly graduate students participating in Phase 1 was 597/268; 522/191 in Phase 2; 939/292 in Phase 3; 575/852 in Phase 4; and 601/1,097 in Phase 5. During this project period, 3,488 papers were published in major scientific journals. Of these, 2,320 papers were published in domestic journals and 1,168 papers were published in Science Citation Index(SCI) journals. During the project period, a total of 83.3 billion won (annual average of 5.5 billion won) or approximately US $75 million (annual average of US $5 million) was invested in investigating indigenous species and collecting specimens. This project was a large-scale research study led by the Korean government. It is considered to be a successful example of Korea's compressed development as it attracted almost all of the taxonomists in Korea and made remarkable achievements with a massive budget in a short time. The results from this project led to the National List of Species of Korea, where all species were organized by taxonomic classification. Information regarding the National List of Species of Korea is available to experts, students, and the general public (https://species.nibr.go.kr/index.do). The information, including descriptions, DNA sequences, habitats, distributions, ecological aspects, images, and multimedia, has been digitized, making contributions to scientific advancement in research fields such as phylogenetics and evolution. The species information also serves as a basis for projects aimed at species distribution and biological monitoring such as climate-sensitive biological indicator species. Moreover, the species information helps bio-industries search for useful biological resources. The most meaningful achievement of this project can be in providing support for nurturing young taxonomists like graduate students. This project has continued for the past 15 years and is still ongoing. Efforts to address issues, including species misidentification and invalid synonyms, still have to be made to enhance taxonomic research. Research needs to be conducted to investigate another 50,000 species out of the estimated 100,000 indigenous species on the Korean Peninsula.