• Title/Summary/Keyword: sequence alignment

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Molecular Cloning and Characterization of Bovine CYP26A1 Promoter (소 CYP26A1 유전자 프로모터의 molecular cloning 및 특성)

  • Kwak, Inseok
    • Journal of Life Science
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    • v.26 no.1
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    • pp.42-49
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    • 2016
  • The retinoic acid (RA) plays an important role in the growth and development of many cells, and bioactive RA concentration is regulated by several enzymes, including CYP26A1. The expression of the CYP26A1 gene is regulated by RA, and the CYP26A1 gene is one of the candidates for RA-responsive genes. Although CYP26A1 genes are cloned from several animals, cloning of the CYP26A1 gene from cows has not been reported yet. The promoter region of CYP26A1 from cows was cloned by PCR and analyzed by sequence alignment with human and mouse CYP26A1. The RA-responsive element (RARE), DR-5 (ttggg), was located in this region and was perfectly conserved. The promoter region of bovine CYP26A1, which contains DR-5, was ligated to the luciferase reporter gene on transient transfection assays. The expression of CYP26A1-Luc promoter was activated by ATRA treatment in lung-derived mtCC cells. Co-transfection with RAR-α or -β with ATRA significantly activates the expression of CYP26A1-Luc promoter; however, it was less effective with either RAR-γ or RXR-γ. In addition, the endogenous gene expressions measured by Q-RT-PCR in mtCC cells were not significantly affected by ATRA treatment for 2 days; however, the expression of the endogenous CYP26A1 gene was diminished sharply at day 3 with ATRA treatment. In conclusion, the promoter region of bovine CYP26A1 contains conserved DR-5 RARE, which functions as a binding site for RAR-α or -β, and it is involved in the regulation of CYP26A1 gene expression and the control of RA signaling in mtCC cells.

Phylogenetic Study of Genus Haliotis In Korea by Internal Transcribed Spacer Sequence (ITS) (ITS에 의한 한국내 전복 속 분류군의 유전적 계통분류학적 연구)

  • Huh, Man-Kyu;Kim, Jung-Ho;Moon, Du-Ho
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1003-1008
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    • 2009
  • Abalone (genus Haliotis) is a woody species with a long life span that is primarily distributed throughout the world, including Asia. This species is regarded as a very important marine gastropod mollusk in Korea and China, and also in food industries around the world. We evaluated a representative sample of the five species with nuclear ribosomal DNA internal transcribed spacer sequences (ITS) to estimate genetic relationships within the genus. Aligned nucleotide sequences of the length of the 5.8S subunit of all taxa of Haliotis were found to constant of 160 bp nucleotides. However, aligned nucleotide sequences of the length of ITS1 were varied within genus Haliotis, varying from 272 in H. diversicolor aquatilis to 292 in H. discus hannai. Aligned nucleotide sequences of the length of ITS2, especially, vary from 722 in H. diversicolor aquatilis to 752 in H. sieboldii. Total alignment length is 763 positions, of which 78 are parsimony-informative, 57 variable but parsimony-uninformative, and 459 constant characters. H. discus hannai was similar to H. discus, while H. diversicolor aquatilis was more distinct. ITS analysis may be useful in germ-plasm classification several taxa of genus Haliotis.

Construction of web-based Database for Haliotis SNP (웹기반 전복류 (Haliotis) SNP 데이터베이스 구축)

  • Jeong, Ji-Eun;Lee, Jae-Bong;Kang, Se-Won;Baek, Moon-Ki;Han, Yeon-Soo;Choi, Tae-Jin;Kang, Jung-Ha;Lee, Yong-Seok
    • The Korean Journal of Malacology
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    • v.26 no.2
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    • pp.185-188
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    • 2010
  • The Web-based the genus Haliotis SNP database was constructed on the basis of Intel Server Platform ZSS130 dual Xeon 3.2 GHz cpu and Linux-based (Cent OS) operating system. Haliotis related sequences (2,830 nucleotide sequences, 9,102 EST sequences) were downloaded through NCBI taxonomy browser. In order to eliminate vector sequences, we conducted vector masking step using cross match software with vector sequence database. In addition, poly-A tails were removed using Trimmest software from EMBOSS package. The processed sequences were clustered and assembled by TGICL package (TIGR tools) equipped with CAP3 software. A web-based interface (Haliotis SNP Database, http://www.haliotis.or.kr) was developed to enable optimal use of the clustered assemblies. The Clustering Res. menu shows the contig sequences from the clustering, the alignment results and sequences from each cluster. And also we can compare any sequences with Haliotis related sequences in BLAST menu. The search menu is equipped with its own search engine so that it is possible to search all of the information in the database using the name of a gene, accession number and/or species name. Taken together, the Web-based SNP database for Haliotis will be valuable to develop SNPs of Haliotis in the future.

Phylogenetic Study of Genus Sorbus in Korea by Internal Transcribed Spacer Sequence (ITS) (ITS에 의한 한국내 마가목 속 분류군의 유전적 계통분류학적 연구)

  • Huh, Man-Kyu;Kim, Sea-Hyun;Park, So-Hye
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1610-1615
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    • 2007
  • Genus Sorbus is a long lived woody species that is primarily distributed throughout Asia and Europe. This species is regarded as very important herbal medicines in Korea and China. Sorbus commixta is primarily distributed throughout Europe. We evaluated a representative sample of the four taxa with nuclear ribosomal DNA internal transcribed spacer sequences (ITS) to estimate genetic relationships within genus. Aligned nucleotide sequences of the length of ITS1 were nearly constant within genus Sorbus varying from 219 in S. aucuparia to 218 in the rest species. Especially, the 5.8S subunit of all taxa of Sorbus was found to constant of 165 bp nucleotides. However, aligned nucleotide sequences of the length of ITS2 vary from 240 in S. sambucifolia var. pseudogrcilisto 245 in S. aucuparia. Total alignment length is 629 positions, of which 35 are parsimony-informative, 32 variable but parsimony-uninformative, and 552 constant characters. The base furtherance showed the difference to the by a total taxon: an average A and T are 17.7% and G and C are 30.4%, 34.2%, respectively. All the four taxa beginning with conserved base paired triplets emerging from single strand regions (domain I). Noteworthy, in the RNA secondary structure proposed for the three Korean Sorbus taxa RNA transcript ITS2, which shows a remarkedly well-conserved folding (domain II). When compared to the European Sorbus (S. aucuparia) of ITS2. ITS analysis may be useful in germ-plasm classification several taxa of genus Sorbus.

Novel reassortant 2.3.4.4B H5N6 highly pathogenic avian influenza viruses circulating among wild, domestic birds in Xinjiang, Northwest China

  • Zhang, Qian;Mei, Xindi;Zhang, Cheng;Li, Juan;Chang, Nana;Aji, Dilihuma;Shi, Weifeng;Bi, Yuhai;Ma, Zhenghai
    • Journal of Veterinary Science
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    • v.22 no.4
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    • pp.43.1-43.10
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    • 2021
  • Background: The H5 avian influenza viruses (AIVs) of clade 2.3.4.4 circulate in wild and domestic birds worldwide. In 2017, nine strains of H5N6 AIVs were isolated from aquatic poultry in Xinjiang, Northwest China. Objectives: This study aimed to analyze the origin, reassortment, and mutations of the AIV isolates. Methods: AIVs were isolated from oropharyngeal and cloacal swabs of poultry. Identification was accomplished by inoculating isolates into embryonated chicken eggs and performing hemagglutination tests and reverse transcription polymerase chain reaction (RT-PCR). The viral genomes were amplified with RT-PCR and then sequenced. The sequence alignment, phylogenetic, and molecular characteristic analyses were performed by using bioinformatic software. Results: Nine isolates originated from the same ancestor. The viral HA gene belonged to clade 2.3.4.4B, while the NA gene had a close phylogenetic relationship with the 2.3.4.4C H5N6 highly pathogenic avian influenza viruses (HPAIVs) isolated from shoveler ducks in Ningxia in 2015. The NP gene was grouped into an independent subcluster within the 2.3.4.4B H5N8 AIVs, and the remaining six genes all had close phylogenetic relationships with the 2.3.4.4B H5N8 HPAIVs isolated from the wild birds in China, Egypt, Uganda, Cameroon, and India in 2016-2017, Multiple basic amino acid residues associated with HPAIVs were located adjacent to the cleavage site of the HA protein. The nine isolates comprised reassortant 2.3.4.4B HPAIVs originating from 2.3.4.4B H5N8 and 2.3.4.4C H5N6 viruses in wild birds. Conclusions: These results suggest that the Northern Tianshan Mountain wetlands in Xinjiang may have a key role in AIVs disseminating from Central China to the Eurasian continent and East African.

Development of Selectable Marker of High Oleate Trait in Peanut (Arachis hypogaea L.) (땅콩에서 고 올레인산 형질관련 분자마커의 선발)

  • Yang, Kiwoung;Pae, Suk-Bok;Park, Chang-Hwan;Lee, Myoung Hee;Jung, Chan-Sik;Son, Jeong-Hee;Park, Keum-Yong
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.507-514
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    • 2010
  • Peanut(Arachis hypogaea L.) is one of the major oilseed crops. The peanut oil consists of palmitic, oleic and linoleic acids, which are present at levels of 10%, 36-67% and 15-43%, respectively. High oleate mutant of peanut F435 contains 80% oleate and as little as 2% linoleate in seed oil. Previous study indicated that delta 12 fatty acid desaturase is a major enzyme controlling the oleate content in seeds of oilseed crops. F435 sequence alignment of their coding regions disclosed that an extra A(adenine) was inserted at the position +2,823 bp of delta 12 fatty acid desaturase gene. This study was to develop molecular marker (SNP marker) co-segregating with the high oleate trait. Chopyeong ${\times}$ F435 $F_2$ 41 population were investigated using molecular marker and fatty acid assay (NIR and gas chromatography). Finally, this marker segregates Chopyeong type 26 lines, heterotype 9 lines and F435 type 6 lines. These results in our study suggested that SNP marker conform fatty acid assay.

Analysis of Spatial Trip Regularity using Trajectory Data in Urban Areas (도시부 경로자료를 이용한 통행의 공간적 규칙성 분석)

  • Lee, Su jin;Jang, Ki tae
    • The Journal of The Korea Institute of Intelligent Transport Systems
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    • v.17 no.6
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    • pp.96-110
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    • 2018
  • As the development of ICT has made it easier to collect various traffic information, research on creating new traffic attributes is drawing attention. Estimation and forecasts of demand and traffic volume are one of the main indicators that are essential to traffic operation, assuming that the traffic pattern at a particular node or link is repeated. Traditionally, a survey method was used to demonstrate this similarity on trip behavior. However, the method was limited to achieving high accuracy with high costs and responses that relied on the respondents' memory. Recently, as traffic data has become easier to gather through ETC system, smart card, studies are performed to identify the regularity of trip in various ways. In, this study, route-level trip data collected in Daegu metropolitan city were analyzed to confirm that individual traveler forms a spatially similar trip chain over several days. For this purpose, we newly define the concept of spatial trip regularity and assess the spatial difference between daily trip chains using the sequence alignment algorithm, Dynamic Time Warping. In addition, we will discuss the applications as the indicators of fixed traffic demand and transportation services.

Microbial Forensics: Comparison of MLVA Results According to NGS Methods, and Forensic DNA Analysis Using MLVA (미생물법의학: 차세대염기서열분석 방법에 따른 MLVA 결과 비교 및 이를 활용한 DNA 감식)

  • Hyeongseok Yun;Seungho Lee;Seunghyun Lim;Daesang Lee;Sehun Gu;Jungeun Kim;Juhwan Jeong;Seongjoo Kim;Gyeunghaeng Hur;Donghyun Song
    • Journal of the Korea Institute of Military Science and Technology
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    • v.27 no.4
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    • pp.507-515
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    • 2024
  • Microbial forensics is a scientific discipline for analyzing evidence related to biological crimes by identifying the origin of microorganisms. Multiple locus variable number tandem repeat analysis(MLVA) is one of the microbiological analysis methods used to specify subtypes within a species based on the number of tandem repeat in the genome, and advances in next generation sequencing(NGS) technology have enabled in silico anlysis of full-length whole genome sequences. In this paper, we analyzed unknown samples provided by Robert Koch Institute(RKI) through The United Nations Secretary-General's Mechanism(UNSGM)'s external quality assessment exercise(EQAE) project, which we officially participated in 2023. We confirmed that the 3 unknown samples were B. anthracis through nucleic acid isolation and genetic sequence analysis studies. MLVA results on 32 loci of B. anthracis were analysed by using genome sequences obtained from NGS(NextSeq and MinION) and Sanger sequencing. The MLVA typing using short-reads based NGS platform(NextSeq) showed a high probability of causing assembly error when a size of the tandem repeats was grater than 200 bp, while long-reads based NGS platform(MinION) showed higher accuracy than NextSeq, although insertion and deletion was observed. We also showed hybrid assembly can correct most indel error caused by MinION. Based on the MLVA results, genetic identification was performed compared to the 2,975 published MLVA databases of B. anthracis, and MLVA results of 10 strains were identical with 3 unkonwn samples. As a result of whole genome alignment of the 10 strains and 3 unknown samples, all samples were identified as B. anthracis strain A4564 which is associated with injectional anthrax isolates in heroin users.

Molecular Phylogenetic Study of the Endangered Land Snail Satsuma myomphala Based on Metallothionein Gene. (Metallothionein 유전자를 기초로 한 멸종위기 육상 달팽이 Satsuma myomphala (거제외줄달팽이) 의 분자계통학적 연구)

  • Sang, Min Kyu;Kang, Se Won;Hwang, Hee-Ju;Chung, Jong Min;Song, Dae Kwon;Min, Hye Rin;Park, Jie Eun;Ha, Hee Cheol;Lee, Hyun Jun;Hong, Chan Eui;Ahn, Young Mo;Park, So Young;Park, Young-Su;Park, Hong Seog;Han, Yeon Soo;Lee, Jun Sang;Lee, Yong Seok
    • The Korean Journal of Malacology
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    • v.32 no.4
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    • pp.263-268
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    • 2016
  • Metallothionein (MT) family of metal-binding proteins are involved in maintaining homeostasis and heavy metal poisoning. Recently, MT has been considered as a biomarker that can identify a particular species, very similar to the use of cytochrome oxidase I (COI) gene. Satsuma myomphala species of land snails have been reported from North-East Asia, including South Korea and Japan. In particular, the land snail species have been known from only a limited area of Geoje Island, Gyeongsangnam-do province of South Korea. Genetic studies of S. myomphala has been limited with only 6 nucleotide, 2 protein registered on the NCBI server. For elucidating the genetic information of S. myomphala, we conducted RNA sequencing analysis using Illumina HiSeq 2500 next-generation platform. We screened the MT gene from the RNA-Seq database to confirm the molecular phylogenetic relationship. After sequencing, the de novo analysis and clustering generated 103,774 unigenes. After annotation against PANM database using BLAST program, we obtained MT sequence of 74 amino acid residues containing the coding region of 222 bp. Based on this sequence, we found about 53 sequences using the BLAST program in NCBI nr database. Using ClustalX alignment, Maximum-Likehood Tree of MEGA program, we confirmed the molecular phylogenetic relationships that showed similarity with mollusks such as Helix pomatia and H. aspersa, Megathura crenulata.

Molecular Characterization of Cinnamate 4-Hydroxylase gene in Red Hot Pepper (Capsicum annuum L.) (고추에서 분리한 Cinnamate 4-Hydroxylase 유전자의 분자생물학적 특성)

  • Kim Kye-Won;Ha Sun-Hwa;Cho Kang-Jin;Kim Eun-Ju;Lee Min-Kyung;Yu Jae-Ju;Kim Jong-Guk;Lee Shin-Woo
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.167-173
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    • 2005
  • Three different cDNAS for cinnamate 4-hydroxylase (C4H) which are involved in the second step of the general phenylpropanoid pathway were isolated and designated as pc4h1 (1,755 bp), pc4h2 (1,655 bp), and pc4h3 (1,316 bp), respectively. The nucleotide sequence analysis revealed that both pc4h1 and pc4h2 clones encode polypeptides of 505 amino acids frame but pc4h3 clone was truncated at the 5'-end of coding region. The alignment of the deduced amino acid sequences showed that PC4H1 and PC4H2 are highly homologous (95.8% identical) with each other and contain three conserved domains which are typical in cytochrome P450 monooxygenase: proline-rich region, threonine-containing binding pocket for the oxygen molecule, and heme binding region. In addition, result of the phylogenic tree analysis revealed that both pepper C4Hs belong to Class 1. pc4h2 transcription was strongly induced in wounded fruit (400%) and root (200%) relative to its very low basal level but not in leaf or stem tissue. In case of pc4h1, the basal level of transcription was higher than pc4h2 but induction by wounding was lower in fruit and root while leaf and stem tissues did not respond to wounding. The basal level of pc4h3 transcripts was not, if any, detectable and response to wounding was not observed.