• Title/Summary/Keyword: Gene assembly

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Investigation of lifespan related genes of the silkmoth, Bombyx mori L (누에나방 수명관련 특이발현 유전자 탐색)

  • Choi, Kwang-Ho;Goo, Tae-Won;Kim, Seong-Ryul;Kim, Sung-Wan;Kang, Seok-Woo;Kang, Pil-Don
    • Journal of Sericultural and Entomological Science
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    • v.51 no.2
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    • pp.211-217
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    • 2013
  • In general, the mean silkmoth lifespan is around 8 days for female and 5 days for male. But, the duration of J037 strain's lifespan is remarkably long in both sexes. On the contrary, the Daizo(sdi) strain has a remarkably short lifetime. The differences in adult lifetime among various silkworm strains has been suggested that the adult lifetime may be genetically controlled. In this experiment, using J037 and Daizo strains we investigated genetic factors related to the adult lifetime of silkworm. We constructed the full-length cDNA library from the adult male of the J037 strain. A total of 2,688 clones were randomly selected, and we performed a differential display hybridization with cDNA probes generated from J037 and Daizo adult males. In conclusion, 193 clones were identified as differential expressed genes, and 154 unique genes were generated after the assembly of 193 clones. Of the 154 unique genes, the most abundant genes were cytochrome oxidase subunit-1 gene(9 times) and unknown(clone ID; 1-50) gene(5 times). The functional groups of these unique genes with matches in the AmiGo database were constructed according to their putative molecular functions. Among thirteen functional categories, the largest group was unclassified protein(24%). In addition, we analyzed the nucleotide and deduced amino acid sequences of the most highly occurred gene(1-50, EF434397), which consisted of 240 amino acids. However, it is confirmed yet that these genes really have an affected on the silkworms longevity. Further studies on these molecules biological roles will give us well-fined information about mechanisms of insect aging and/or scenesence.

Crystal Structure of the Regulatory Domain of AphB from Vibrio vulnificus, a Virulence Gene Regulator

  • Park, Nohra;Song, Saemee;Choi, Garam;Jang, Kyung Ku;Jo, Inseong;Choi, Sang Ho;Ha, Nam-Chul
    • Molecules and Cells
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    • v.40 no.4
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    • pp.299-306
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    • 2017
  • The transcriptional activator AphB has been implicated in acid resistance and pathogenesis in the food borne pathogens Vibrio vulnificus and Vibrio cholerae. To date, the full-length AphB crystal structure of V. cholerae has been determined and characterized by a tetrameric assembly of AphB consisting of a DNA binding domain and a regulatory domain (RD). Although acidic pH and low oxygen tension might be involved in the activation of AphB, it remains unknown which ligand or stimulus activates AphB at the molecular level. In this study, we determine the crystal structure of the AphB RD from V. vulnificus under aerobic conditions without modification at the conserved cysteine residue of the RD, even in the presence of the oxidizing agent cumene hydroperoxide. A cysteine to serine amino acid residue mutant RD protein further confirmed that the cysteine residue is not involved in sensing oxidative stress in vitro. Interestingly, an unidentified small molecule was observed in the inter-subdomain cavity in the RD when the crystal was incubated with cumene hydroperoxide molecules, suggesting a new ligand-binding site. In addition, we confirmed the role of AphB in acid tolerance by observing an aphB-dependent increase in cadC transcript level when V. vulnificus was exposed to acidic pH. Our study contributes to the understanding of the AphB molecular mechanism in the process of recognizing the host environment.

Current biotechnology for the increase of vegetable oil yield in transgenic plants (식물 지방산 생산량의 증진을 위한 생명공학 연구현황)

  • Lee, Kyeong-Ryeol;Choi, Yun-Jung;Kim, Sun-Hee;Roh, Kyung-Hee;Kim, Jong-Bum;Kim, Hyun-Uk
    • Journal of Plant Biotechnology
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    • v.38 no.4
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    • pp.241-250
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    • 2011
  • The most part of vegetable oils is accumulated as storage lipid, triacylglycerol (TAG) in seed and used as energy source when seed is germinated. It is also used as essential fatty acids and energy source for human and animal. Recently, vegetable oils have been more and more an important resource because of the increasing demand of vegetable oils for cooking and industrial uses for bio-diesel and industrial feedstock. In order to increase vegetable oils using biotechnology, over-expressing or repressing the regulatory genes involved in the flow of carbon into lipid biosynthesis is critical during seed development. In this review, we described candidate genes may influence oil amount and investigate their potential for oil increase. Genes involved in the regulation from biosynthesis of fatty acids to the accumulation oils in seed can be classified as follows: First, genes play a role for synthesis precursor molecules for TAG. Second, genes participate in fatty acid biosynthesis and TAG assembly. Lastly, genes encodes transcription factors involved in seed maturation and accumulation of seed oil. Because factors/genes determining oil quantity in seed is complex as mentioned, recently regulation of transcription factors is being considered more favorable approach than manipulate multiple genes for increasing oil in transgenic plants. However, it should be figured out the problem that bad agricultural traits induced by the overexpression of transcription factor gene.

Transcriptome profiling and comparative analysis of Panax ginseng adventitious roots

  • Jayakodi, Murukarthick;Lee, Sang-Choon;Park, Hyun-Seung;Jang, Woojong;Lee, Yun Sun;Choi, Beom-Soon;Nah, Gyoung Ju;Kim, Do-Soon;Natesan, Senthil;Sun, Chao;Yang, Tae-Jin
    • Journal of Ginseng Research
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    • v.38 no.4
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    • pp.278-288
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    • 2014
  • Background: Panax ginseng Meyer is a traditional medicinal plant famous for its strong therapeutic effects and serves as an important herbal medicine. To understand and manipulate genes involved in secondary metabolic pathways including ginsenosides, transcriptome profiling of P. ginseng is essential. Methods: RNA-seq analysis of adventitious roots of two P. ginseng cultivars, Chunpoong (CP) and Cheongsun (CS), was performed using the Illumina HiSeq platform. After transcripts were assembled, expression profiling was performed. Results: Assemblies were generated from ~85 million and ~77 million high-quality reads from CP and CS cultivars, respectively. A total of 35,527 and 27,716 transcripts were obtained from the CP and CS assemblies, respectively. Annotation of the transcriptomes showed that approximately 90% of the transcripts had significant matches in public databases.We identified several candidate genes involved in ginsenoside biosynthesis. In addition, a large number of transcripts (17%) with different gene ontology designations were uniquely detected in adventitious roots compared to normal ginseng roots. Conclusion: This study will provide a comprehensive insight into the transcriptome of ginseng adventitious roots, and a way for successful transcriptome analysis and profiling of resource plants with less genomic information. The transcriptome profiling data generated in this study are available in our newly created adventitious root transcriptome database (http://im-crop.snu.ac.kr/transdb/index.php) for public use.

Development of Selectable Vector Plasmid in Bacteriophage P2-P4 System and Its Stability (박테리오파지 P2-P4 시스템을 위한 벡터 플라스미드 개발과 안정성)

  • Kim, Kyoung-Jin
    • Korean Journal of Microbiology
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    • v.34 no.4
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    • pp.236-242
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    • 1998
  • While bacteriophage P2-P4 system is very useful experimental tool for the study of viral capsid assembly, there is no useful plasmid vector for the DNA manipulation in bacteriophage P2-P4 system. In this study, a new vector plasmid, P4 ash8 (sid71) kmr, was constructed by swapping the non-essential region of P4 DNA for kanamycin resistance(kmr) gene cassette of plasmid pUC4-K. P4 ash8 sid71 was starting material for the construction, since it tends to be maintained as a plasmid in the absence of the helper phage. The total size of this chimera was designed to be packaged into P4 or P2 size heads with induction by P2 infection. The conversion of plasmid P4 ash8 (sid71) kmr to bacteriophage was proved by burst size determination experiment and CsCl buoyant equilibrium density gradient experiment. Integrase destructed P4 derivative, P4 ash8 sid71 kmr intS, was able to be constructed easily by in vitro DNA manipulation of P4 ash8 sid71 kmr. The plasmid stability experiment with P4 ash8 sid71 kmr if/tS showed that the integrase of P4 affects the stable maintenance of plasmid P4 state.

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The Association of Long Noncoding RNA LOC105372577 with Endoplasmic Reticulum Protein 29 Expression: A Genome-wide Association Study (ERp29 유전자 발현과 관련된 long noncoding RNA LOC105372577의 전장 유전체 연관성 분석)

  • Lee, Soyeon;Kwon, Kiang;Ko, Younghwa;Kwon, O-Yu
    • Journal of Life Science
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    • v.31 no.6
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    • pp.568-573
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    • 2021
  • This study identified genomic factors associated with endoplasmic reticulum protein (ERp)29 gene expression in a genome-wide association study (GWAS) of genetic variants, including single-nucleotide polymorphisms (SNPs). In total, 373 European genes from the 1000 Genomes Project were analyzed. SNPs with an allelic frequency of less than or more than 5% were removed, resulting in 5,913,563 SNPs including in the analysis. The following expression quantitative trait loci (eQTL) from the long noncoding RNA LOC105372577 were strongly associated with ERp29 expression: rs6138266 (p<4.172e10), rs62193420 (p<1.173e10), and rs6138267 (p<2.041e10). These were strongly expressed in the testis and in the brain. The three eQTL were identified through a transcriptome-wide association study (TWAS) and showed a significant association with ERp29 and osteosarcoma amplified 9 (OS9) expression. Upstream sequences of rs6138266 were recognized by ChIP-seq data, while HaploReg was used to demonstrate how its regulatory DNA binds upstream of transcription factor 1 (USF1). There were no changes in the expression of OS9 or USF1 following ER stress.

Comparative transcriptome and metabolome analyses of four Panax species explore the dynamics of metabolite biosynthesis

  • Hyunjin, Koo;Yun Sun, Lee;Van Binh, Nguyen;Vo Ngoc Linh, Giang;Hyun Jo, Koo;Hyun-Seung, Park;Padmanaban, Mohanan;Young Hun, Song;Byeol, Ryu;Kyo Bin, Kang;Sang Hyun, Sung;Tae-Jin, Yang
    • Journal of Ginseng Research
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    • v.47 no.1
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    • pp.44-53
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    • 2023
  • Background: The genus Panax in the Araliaceae family has been used as traditional medicinal plants worldwide and is known to biosynthesize ginsenosides and phytosterols. However, genetic variation between Panax species has influenced their biosynthetic pathways is not fully understood. Methods: Simultaneous analysis of transcriptomes and metabolomes obtained from adventitious roots of two tetraploid species (Panax ginseng and P. quinquefolius) and two diploid species (P. notoginseng and P. vietnamensis) revealed the diversity of their metabolites and related gene expression profiles. Results: The transcriptome analysis showed that 2,3-OXIDOSQUALENE CYCLASEs (OSCs) involved in phytosterol biosynthesis are upregulated in the diploid species, while the expression of OSCs contributing to ginsenoside biosynthesis is higher in the tetraploid species. In agreement with these results, the contents of dammarenediol-type ginsenosides were higher in the tetraploid species relative to the diploid species. Conclusion: These results suggest that a whole-genome duplication event has influenced the triterpene biosynthesis pathway in tetraploid Panax species during their evolution or ecological adaptation. This study provides a basis for further efforts to explore the genetic variation of the Panax genus.

Partial Sequencing and Characterization of Porcine DNA Methyltransferase I cDNA

  • Lee, Y.Y.;Kim, M.S.;Park, J.J.;H.Y. Kang;Y.M. Chang;Yoon, J.T.;K.S. Min
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.84-84
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    • 2003
  • DNA methylation is involved in epigenetic processes such as X-chromosome inactivation, imprinting and silencing of transposons. DNA methylation is a highly plastic and critical component of mammalian development The DNA methyltransferases (Dnmts) are responsible for the generation of genomic methylation patterns, which lead to transcriptional silencing. The maintenance DNA methyltransferase enzyme, Dnmt 1, and the de novo methyltransferase, Dnmt3a and Dnmt3b, are indispensable for development because mice homozygous for the targeted disruption of any of these genes are not viable. The occurrence of DNA methylation is not random, and it can result in gene silencing The mechanisms underlying these processes are poorly understood. It is well established that DNA methylation and histone deacetylation operate along a common mechanistic pathway to repress transcription through the action of methyl-binding domain proteins (MBDs), which are components of, or recruit, histone deacetylase (HDAC) complexes to methylated DNA. As a basis for future studies on the role of the DNA-methyl-transferase in porcine development, we have isolated and characterized a partial cDNA coding for the porcine Dnmt1. Total RNA of testis, lung and ovary was isolated with TRlzol according to the manufacture's specifications. 5 ug of total RNA was reverse transcribed with Super Script II in the presence of porcine Dnmt 1 specific primers. Standard PCRs were performed in a total volume of 50 ul with cDNA as template. Two DNA fragmenets in different position were produced about 700bp, 1500bp and were cloned into pCR II-TOPO according to the manufacture's specification. Assembly of all sequences resulted in a cDNA from 158bp of 5'to 4861bp of 3'compare with the known human maintenance methyltransferase. Now, we are cloning the unknown Dnmt 1 region by 5'-RACE method and expression of Dnmt 1 in tissues from adult porcine animals.

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The Philippines Coconut Genomics Initiatives: Updates and Opportunities for Capacity Building and Genomics Research Collaboration

  • Hayde Flandez-Galvez;Darlon V. Lantican;Anand Noel C. Manohar;Maria Luz J. Sison;Roanne R. Gardoce;Barbara L. Caoili;Alma O. Canama-Salinas;Melvin P. Dancel;Romnick A. Latina;Cris Q. Cortaga;Don Serville R. Reynoso;Michelle S. Guerrero;Susan M. Rivera;Ernesto E. Emmanuel;Cristeta Cueto;Consorcia E. Reano;Ramon L. Rivera;Don Emanuel M. Cardona;Edward Cedrick J. Fernandez ;Robert Patrick M. Cabangbang;Maria Salve C. Vasquez;Jomari C. Domingo;Reina Esther S. Caro;Alissa Carol M. Ibarra;Frenzee Kroeizha L. Pammit;Jen Daine L. Nocum;Angelica Kate G. Gumpal;Jesmar Cagayan;Ronilo M. Bajaro;Joseph P. Lagman;Cynthia R. Gulay;Noe Fernandez-Pozo;Susan R. Strickler;Lukas A. Mueller
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.30-30
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    • 2022
  • Philippines is the second world supplier of coconut by-products. As its first major genomics project, the Philippine Genome Center program for Agriculture (PGC-Agriculture) took the challenge to sequence and assemble the whole coconut genome. The project aims to provide advance genetics tools for our collaborating coconut researchers while taking the opportunity to initiate local capacity. Combination of different NGS platforms was explored and the Philippine 'Catigan Green Dwarf' (CATD) variety was selected with the breeders to be the crop's reference genome. A high quality genome assembly of CATD was generated and used to characterize important genes of coconut towards the development of resilient and outstanding varieties especially for added high-value traits. The talk will present the significant results of the project as published in various papers including the first report of whole genome sequence of a dwarf coconut variety. Updates will include the challenges hurdled and specific applications such as gene mining for host insect resistance and screening for least damaged coconuts (thus potentially insect resistant varieties). Genome-wide DNA markers as published and genes related to coconut oil qualitative/quantitative traits will also be presented, including initial molecular/biochemical studies that support nutritional and medicinal claims. A web-based genome database is currently built for ease access and wider utility of these genomics tools. Indeed, a major milestone accomplished by the coconut genomics research team, which was facilitated with the all-out government support and strong collaboration among multidisciplinary experts and partnership with advance research institutes.

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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.