• Title/Summary/Keyword: Genome Length

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Identification and Expression of Retroviral Envelope Polyprotein in the Dogfish Squalus mitsukurii

  • Kim, Soo Cheol;Sumi, Kanij Rukshana;Choe, Myeong Rak;Kho, Kang Hee
    • Journal of Marine Life Science
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    • v.1 no.2
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    • pp.88-94
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    • 2016
  • Determining the infection history of living organisms is essential for understanding the evolution of infection agents with their host, particularly for key aspects such as immunity. Viruses, which can spread between individuals and often cause disease, have been widely examined. The increasing availability of fish genome sequences has provided specific insights into the diversity and host distribution of retroviruses in fish. The shortspine spurdog (Squalus mitsukurii) is an important elasmobranch species; this medium-sized dogfish typically lives at depths of 100~500 m. However, the retroviral envelope polyprotein in dogfish has not been examined. Thus, the aim of the present study was to identify and analyze the retroviral envelope polyprotein in various tissues of dogfish. The 1334-base pair full-length novel cDNA of dogfish envelope polyprotein (dEnv) was obtained by 3' and 5'-rapid amplification of cDNA end analysis from S. mitsukurii. The open reading frame showed a complete coding sequence of 815 base pairs with a deduced peptide sequence of 183 amino acids that exhibited 34~50% identity with other fish and bird species. It was also expressed according to reverse transcription and real-time polymerase chain reaction in the kidney, liver, intestine, and lung, but not in the gill. This distribution can be assessed by identifying and analyzing endogenous retroviruses in fish, which consists of three main genes: gag, pol and env. Dogfish envelope polyprotein sequence is likely important in evolution and induces rearrangements, altering the regulatory and coding sequences. This is the first report of the identification and molecular characterization of retroviral envelope polyprotein in various tissues of S. mitsukurii.

Lack of association between the VEGFA gene polymorphisms and preterm birth in Korean women

  • Yue Shi;Hyung Jun Kim;Seong Yong Kim;Ga Eun Kim;Han Jun Jin
    • Genomics & Informatics
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    • v.21 no.3
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    • pp.29.1-29.9
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    • 2023
  • Preterm birth (PTB), a pregnancy-related disease, is defined as a birth before 37 weeks of gestation. It is a major cause of maternal mortality and morbidity worldwide, and its incidence rate is steadily increasing. Various genetic factors can contribute to the etiology of PTB. Vascular endothelial growth factor A (VEGFA) gene is an important angiogenic gene and its polymorphisms have been reported to be associated with PTB development. Therefore, we conducted a case-control study to evaluate the association between VEGFA rs699947, rs2010963, and rs3025039 polymorphisms and PTB in Korean women. A total of 271 subjects (116 patients with PTB and 155 women at ≥38 weeks of gestation) were analyzed in this study. The genotyping of VEGFA gene polymorphisms was performed using polymerase chain reaction- restriction fragment length polymorphism. No significant association between the patients with PTB and the control groups was confirmed. In the combination analysis, we found a significant association between PTB and VEGFA rs699947 CC-rs2010963 GG-rs3025039 CC combination (odds ratio, 3.77; 95% confidence interval, 1.091 to 13.032; p = 0.031). The VEGFA rs699947, rs2010963, and rs3025039 polymorphisms might have no genetic association with the pathogenesis of PTB in Korean women. However, the combination analysis indicates the possibility that VEGFA acts in PTB pathophysiology. Therefore, larger sample sets and replication studies are required to further elucidate our findings.

Isolation and Characterization of a Novel Flavonoid 3'-Hydroxylase (F3'H) Gene from a Chrysanthemum (Dendranthema grandiflorum) and Its Gamma-ray Irradiated Mutants (감마선 처리에 의한 스프레이형 국화 화색변이체로부터 Flavonoid 3'-Hydroxylase(F3'H) 유전자의 분리 및 특성 구명)

  • Chung, Sung-Jin;Lee, Geung-Joo;Kim, Jin-Baek;Kim, Dong-Sub;Kim, Sang-Hoon;Kang, Si-Yong
    • Horticultural Science & Technology
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    • v.30 no.2
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    • pp.162-170
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    • 2012
  • The objectives of this study were to isolate and the sequence of novel $F3'H$ gene related to an anthocyanin pathway, and to confirm the expression patterns of the gene involved in the flower color variations of chrysanthemum mutants. In this study, we isolated the full-length cDNAs and the genomic DNAs of an $F3'H$ gene from a wild type (WT) chrysanthemum (cv. Argus) and its three color mutants. The sequence analysis revealed a putative open reading frame of 1,527 bp that encodes a polypeptide of 509 amino acids. Sequence homology ranged from 97% to 99% between 'Argus' and its three color mutants. The sequence analysis from the genomic DNA revealed that the chrysanthemum $DgF3'H$ gene consisted of three exons and two introns spanning a 3,830 bp length. The sizes of the gene for three mutants ranged from a shorter size of 3,828 bp to a longer size of 3,838 bp when compared to the size of WT. The total size of the two introns was 2,157 bp for WT, but those for three color mutants ranged from 2,154 bp to 2,159 bp. A result of an RT-PCR analysis indicated that the color variations of the mutants AM1 and AM2 can be partly explained by the structural modification derived from the sequencial changes in the gene caused by gamma ray. A Southern blot analysis revealed that the $DgF3'H$ gene existing as multiple copies in the chrysanthemum genome. A systemic study will be further needed to provide a genetic mechanism responsible for the color mutation and to uncover any involvement of genetic elements for the expression of the $DgF3'H$ gene for the color variation in chrysanthemum.

Molecular Cloning and Characterization of the Yew Gene Encoding Squalene Synthase from Taxus cuspidata

  • Huang, Zhuoshi;Jiang, Keji;Pi, Yan;Hou, Rong;Liao, Zhihua;Cao, Ying;Han, Xu;Wang, Qian;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.40 no.5
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    • pp.625-635
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    • 2007
  • The enzyme squalene synthase (EC 2.5.1.21) catalyzes a reductive dimerization of two farnesyl diphosphate (FPP) molecules into squalene, a key precursor for the sterol and triterpene biosynthesis. A full-length cDNA encoding squalene synthase (designated as TcSqS) was isolated from Taxus cuspidata, a kind of important medicinal plants producing potent anti-cancer drug, taxol. The full-length cDNA of TcSqS was 1765 bp and contained a 1230 bp open reading frame (ORF) encoding a polypeptide of 409 amino acids. Bioinformatic analysis revealed that the deduced TcSqS protein had high similarity with other plant squalene synthases and a predicted crystal structure similar to other class I isoprenoid biosynthetic enzymes. Southern blot analysis revealed that there was one copy of TcSqS gene in the genome of T. cuspidata. Semi-quantitative RT-PCR analysis and northern blotting analysis showed that TcSqS expressed constitutively in all tested tissues, with the highest expression in roots. The promoter region of TcSqS was also isolated by genomic walking and analysis showed that several cis-acting elements were present in the promoter region. The results of treatment experiments by different signaling components including methyl-jasmonate, salicylic acid and gibberellin revealed that the TcSqS expression level of treated cells had a prominent diversity to that of control, which was consistent with the prediction results of TcSqS promoter region in the PlantCARE database.

Screening and Characterization of LTR Retrotransposons in the genomic DNA of Pleurotus eryngii (큰느타리버섯 유전체내 LTR Retrotransposon 유전자 탐색 및 특성연구)

  • Kim, Sinil;Le, Quy Vang;Kim, Sun-Mi;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.42 no.1
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    • pp.50-56
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    • 2014
  • Transposable elements (TEs) are mobile DNA elements that often cause mutations in genes and alterations in the chromosome structure. In order to identify and characterize transposable elements (TEs) in Pleurotus eryngii, a TE-enriched library was constructed using two sets of TE-specific degenerated primers, which target conserved sequences of RT and RVE domains in fungal LTR retrotransposons. A total of 256 clones were randomly chosen from the library and their insert sequences were determined. Comparative investigation of the insert sequences with those in repeat element database, Repbase, revealed that 71 of them were found to be TE-related fragments with significant similarity to LTR retrotransposons from other species. Among the TE sequences, the 70 TEs were Gypsy-type LTR retrotransposons, including 20 of MarY1 from Tricholoma matsutake, 26 of Gypsy-8_SLL from Serpula lacrymans, and 16 of RMER17D_MM from mouse, whereas a single sequence, Copia-48-PTR, was found as only Copia-type LTR retrotransposon. Southern blot analysis of the HindIII-digested P. eryngii genomic DNA showed that the retrotransposon sequences similar to MarY1 and Gypsy-8_SLL were contained as high as 14 and 18 copies per genome, respectively, whereas other retrotransposons were remained low. Moreover, both of the two Gypsy retrotransposons were expressed in full length mRNA as shown by Northern blot analysis, suggesting that they were functionally active retrotransposons.

Sequence Analysis of Segments 8 and 10 of Rice black-streaked dwarf virus from Maize Plants (옥수수에 발생한 벼검은줄오갈병 바이러스 분절게놈 S8 및 S10 전 염기서열 분석)

  • Lee, Bong Choon;Cho, Sang-Yun;Yoon, Young-Nam;Kang, In Jeong;Kwak, Do Yeon;Shin, Dong Bum;Kang, Hang-Won
    • Research in Plant Disease
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    • v.18 no.4
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    • pp.387-390
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    • 2012
  • Rice black-streaked dwarf virus (RBSDV) was reported to occur on maize plants in Gochang-gun of Jeonllabuk-do region in 2011. The symptoms typically include stunted and deformed leaves. Virus infected plants usually produce poor or no head. RT-PCR analysis of genomic dsRNA extracted from the plant confirmed the infection. Specific primers for full length genome of segments 8 and 10 were used for RNA amplification. Full-length genomes of S8 and S10 were cloned and sequenced. Sequence analysis revealed that the S8 and S10 sequences of the maize isolate were same with rice isolate in size, 1,936 nt and 1,801 nt, respectively. In comparison with rice RBSDV, S8 and S10 showed 94.9-99.6% and 94.1-98.4% sequence identity, respectively. Phylogenetic analysis showed that RBSDV S8 and S10 of maize plants are categorized into the same group as RBSDV of rice plant.

Analysis of Populus cpDNA by Restriction Fragment Length Polymorphism(RFLP) Technique (RFLP기법(技法)을 이용(利用)한 포플러 엽록체(葉綠體) DNA의 분석(分析))

  • Lee, J.S.;Noh, E.W.;Lee, S.K.;Kwon, K.W.
    • Journal of Korean Society of Forest Science
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    • v.83 no.1
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    • pp.20-24
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    • 1994
  • In woody species with a long life span, the studies on inheritance of any trait may be very time consuming and laborious. Chloroplast DNA(cpDNA) has been a valuable tool in such studies since it has several unique features such as limited genome size and cytoplasmic inheritance. In the present study, cpDNAs from five different species of Populus(P. alba, P. glandulosa, P. alba${\times}$P. glandulosa, P. davidiana, and P. nigra), and Nicotiana tabacum were compared with regard to restriction fragment length polymophism. The results showed that cpDNAs among the species were very conserved, although some polymorphisms were observed when the DNAs were digested with restriction enzyme EcoRI or KphI. The other enzymes (Bgl II, and PstI) tested produced identical restriction fragmentation pattern among the species. However, cpDNAs from all the five Populus species showed different restriction fragmentation pattern from that of tobacco with the four restriction enzymes tested. Southern hybridization with tobacco rbcL gene fragment as a probe also produced identical pattern among Populus species. The results indicate that cpDNAs in the genus are very well conserved during evolution.

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Some Characteristics of Melon necrotic spot virus-Me and Resistance Screen to the Virus in Melon Cultivars (멜론괴저반점바이러스-Me의 몇 가지 특성과 멜론 품종의 저항성 선발)

  • Choi, Gug-Seoun;Cho, Jeom-Deog;Chung, Bong-Nam;Cho, In-Sook;Kwon, Soon-Bae
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.254-258
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    • 2010
  • Melon necrotic spot virus (MNSV) is a very destructive disease to melon (Cucumis melo) plants. A MNSV was isolated from melon leaf showing necrotic spot symptoms at the plastic house in Naju, Korea in 2009. The isolate, designated as MNSV-Me, was identified and characterized by biological responses on several host plants, immuno captured RT-PCR and partial nucleotide sequencings of the genome. To evaluate MNSV-Me resistance in melon, thirty-five melon cultivars were mechanically inoculated on the cotyledon of the seedlings with the virus. MNSV-Me produced necrotic spots on the inoculated leaves of the all melon cultivars tested. Twenty-five cultivars were susceptible to the virus and they showed systemic necrotic spots on the leaves and/or necrosis longer than 3 cm in length on the stems within about forty days after inoculation. Five cultivars gave moderate resistance, no symptoms on the upper leaves but necrosis on the stem shorter than 3 cm in length. In an evaluation of MNSV-Me resistance in melon cultivars, 'Elstitan', 'Elsluxery', 'Betalichihage', 'Betalichi' and 'Womderfulhagae 1st' were found to have resistance by showing only faint necrosis on their stems.

Isolation of a Leucoanthocyanidin Dioxygenase (LDOX) Gene from a Spray-type Chrysanthemum (Dendranthema × grandiflorum) and Its Colored Mutants (스프레이형 국화와 화색변이체로부터 Leucoanthocyanidin dioxygenase (LDOX) 유전자의 분리)

  • Chung, Sung-Jin;Lee, Geung-Joo;Lee, Hye-Jung;Kim, Jin-Baek;Kim, Dong-Sub;Kang, Si-Yong
    • Horticultural Science & Technology
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    • v.28 no.5
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    • pp.818-827
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    • 2010
  • A full-length cDNA and genomic DNA of a $leucoanthocyanidin$ $dioxygenase$ ($DgLDOX$) gene was isolated from the petals of chrysanthemum 'Argus', and comparative features of the gene among three flower color mutants derived from a gamma-ray mutagenesis were characterized. The cDNA coding region of the gene was 1068 bp and was translated into 356 amino acids accordingly. The genomic DNA size was 1346 bp for 'Argus', while three mutants revealed ranges of 1363 to 1374 bp. A single intron between two coding exons for the $DgLDOX$ gene was found, of which size was 112 bp for 'Argus', but 128 or 137 bp for three flower color mutants, indicating that a genomic insertion in the intron occurred during the gamma-ray mutagenesis. DNA blot analysis revealed the $DgLDOX$ gene presenting as a single copy in the chrysanthemum genome. The $DgLDOX$ gene was expressed in both 'Argus' of light-pink color and two purple color mutants (AM1 and AM3) but had very weak expression in only white color mutant (AM2). The results demonstrated that variations in the flower color of the mutants might be associated with changes in the amino acid moieties in the coding exons or fragment insertions in the intron of the $DgLDOX$ gene, which potentially resulted in less expression of the gene in the white colored mutant.

Development of Near-isogenic Japonica Rice Lines with Enhanced Resistance to Magnaporthe grisea

  • Kwon, Soon-Wook;Cho, Young-Chan;Kim, Yeon-Gyu;Suh, Jung-Pil;Jeung, Ji-Ung;Roh, Jae-Hwan;Lee, Sang-Kyu;Jeon, Jong-Seong;Yang, Sae-Jun;Lee, Young-Tae
    • Molecules and Cells
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    • v.25 no.3
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    • pp.407-416
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    • 2008
  • Thirteen near-isogenic lines (NILs) of japonica rice were developed via a backcross method using the recurrent parent Chucheong, which is of good eating quality but is susceptible to Magnaporthe grisea, and three blast resistant japonica donors, Seolak, Daeseong and Bongkwang. The agro-morphological traits of these NILs, such as heading date, culm length, and panicle length, were similar to those of Chucheong. In a genome-wide scan using 158 SSR markers, chromosome segments of Chucheong were identified in most polymorphic regions of the 13 NIL plants, and only a few chromosome segments were found to have been substituted by donor alleles. The genetic similarities of the 13 NILs to the recurrent parent Chucheong averaged 0.961, with a range of 0.932-0.984. Analysis of 13 major blast resistance (R) genes in these lines using specific DNA markers showed that each NIL appeared to contain some combination of the four R genes, Pib, Pii, Pik-m and Pita-2, with the first three genes being present in each line. Screening of nine M. grisea isolates revealed that one NIL M7 was resistant to all nine isolates; the remaining NILs were each resistant to between three and seven isolates, except for NIL M106, which was resistant to only two isolates. In a blast nursery experiment, all the NILs proved to be more resistant than Chucheong. These newly developed NILs have potential as commercial rice varieties because of their increased resistance to M. grisea combined with the desirable agronomic traits of Chucheong. They also provide material for studying the genetic basis of blast resistance.