• Title/Summary/Keyword: 3'-UTR nucleotide sequences

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Phylogenetic ANalysis of Hepatitis G Virus by Group-Specific Sequences in the 5-Untranslated Region (5'-UTR 영역의 그룹특이적 염기서열에 의한 HGV의 계통분석)

  • Kim, Pu-Kyung;Park, Sung-Woo;Kim, Chong-Kyung;Baik, Hyung-Suk;Jang, Kyung-Lib
    • Journal of Life Science
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    • v.8 no.3
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    • pp.279-284
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    • 1998
  • The nucleotide sequences of the 5'-untraslated region(5'-UTR) of Hepatitis G virus(HGV) from sera of Korean patients were determines. When compared to the previously reported isolates, the Korean isolates have higher sequence homology with the Japanese isolates indicating the geographic distribution of HGV variants. Interestingly, three discrete regions which are highly conserved among HGV isolates from the same geographical area, thus could be applied to distinguish HGV isolates from the different areas were noticed in the 5'-UTR. Based on the sequences of these group-specific regions, twenty four different HGV isolates could be classified into 5 groups. By using the group-specific regions, inconsistency in HGV typing when based on the different regions of HGV could be solved.

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Cloning of Growth Hormone Complementary DNA from Red-Spotted Grouper (Epinephelus akaara) and Its Expression in E. coli (붉바리(Epinephelus akaara)의 성장호르몬 cDNA의 Cloning과 E. coli에서의 발현)

  • 강거영;송춘복;이제희
    • Journal of Aquaculture
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    • v.16 no.2
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    • pp.110-117
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    • 2003
  • We have cloned and sequenced the cDNA encoding growth hormone (GH) from pituitary poly(A)$^{+}$ RNA of red-spotted grouper (Epinephelus akaara). The cDNA of red-spotted grouper GH is 883 base pairs (bp) consisting of 21 bp of 5'untranslated region (UTR), 615 Up of an open reading frame (ORF) and 247 Up of 3'UTR. The polyadenylation signal, AATAAA, was 20 bp upsteam of polyadenylation site. Based on the nucleotide sequences, the deduced putative polypeptide contains 204 amino acids (aa), representing 17 aa of a signal and 187 aa of a mature polypeptide. The putative GH cDNA encodes a polypeptide with four cysteine residues and only one N-gly- cosylation site. Comparative sequence alignment shows that red-spotted grouper GH exhibits high similarity with its corresponding other Perciformes species GH cDNAs.

Characterization and diagnosis of YMV-K strains infecting Chinese yam(Dioscorea opposita Thunb. cv. Dung-Gun-Ma) in Korea. (oral)

  • D. K. Kang;H. Y. Shin;J. H. Sung;Park, J. H.;M. U. Chang
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.127.2-127
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    • 2003
  • YMV-K strains were purified from D. oppostita Thunb. tv. Dung-Gun-Ma showing mosaic symptom on their leaves. YMV-K strains were filamentous particles of 780nm in length and induced cytoplasmic disorder such as inclusion body formation. Nucleotide and amino acid sequences of 5'-UTR, Pl and CP of YMV-K strains shared 80.8, 64.7 and 98.3% identity respectively to JYMV J1 in the mean value. Purification of YMV-K strains according to JYMV purification method(S. Fuji) was conducted to product antiserum. With antiserum against YMV-K strains, the Various diagnosis methods such as IC-RT-PCR, DIBA, RIPA and indirect-ELISA were used to detect YMV-K strains in Chinese yam plant.

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Sequence Characterization, Expression Profile, Chromosomal Localization and Polymorphism of the Porcine SMPX Gene

  • Guan, H.P.;Fan, B.;Li, K.;Zhu, M.J.;Yerle, M.;Liu, Bang
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.7
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    • pp.931-937
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    • 2006
  • The full-length cDNA of the porcine SMPX gene was obtained by the rapid amplification of cDNA ends (RACE). The nucleotide sequences and the predicted protein sequences share high sequence identity with both human and mouse. The promoter of SMPX was sequenced and then analyzed to find the promoter binding sites. The reverse transcriptase-polymerase chain reaction (RT-PCR) revealed that SMPX has a high level of expression in heart and skeletal muscle, a very low expression in lung and spleen and no expression in liver, kidney, fat and brain. Moreover, SMPX has a differential expression level in skeletal muscle, the expression in 65-day embryos being higher than other stages. The porcine SMPX was mapped to SSCXp24 by using a somatic cell hybrid panel (SCHP) and was found closely linked to SW1903 using the radiation hybrid panel IMpRH. An A/G single nucleotide polymorphism (PCR-RFLP) in the 3'-untranslated region (3'-UTR) was detected in eight breeds. The analysis of allele frequency distribution showed that introduced pig breeds (Duroc and Large White) have a higher frequency of allele A while in the Chinese indigenous pig breeds (Qingping pig, Lantang pig, YushanBlack pig, Large Black-White pig, Small Meishan) have a higher frequencies of allele G. The association analysis using an experimental population (188 pigs), which included two cross-bred groups and three pure-blood groups, suggested that the SNP genotype was associated with intramuscular fat content.

Mutational Analysis of an Essential RNA Stem-loop Structure in a Minimal RNA Substrate Specifically Cleaved by Leishmania RNA Virus 1-4 (LRV1-4) Capsid Endoribonuclease

  • Ro, Youngtae;Patterson, Jean L.
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.239-247
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    • 2003
  • The LRV1-4 capsid protein possesses an endoribonuclease activity that is responsible for the single site-specific cleavage in the 5' untranslated region (UTR) of its own viral RNA genome and the formation of a conserved stem-loop structure (stem-loop IV) in the UTR is essential for the accurate RNA cleavage by the capsid protein. To delineate the nucleotide sequences, which are essential for the correct formation of the stem-loop structure for the accurate RNA cleavage by the viral capsid protein, a wildtype minimal RNA transcript (RNA 5' 249-342) and several synthetic RNA transcripts encoding point-mutations in the stem-loop region were generated in an in vitro transcription system, and used as substrates for the RNA cleavage assay and RNase mapping studies. When the RNA 5' 249-342 transcript was subjected to RNase T1 and A mapping studies, the results showed that the predicted RNA secondary structure in the stem-loop region using FOLD analysis only existed in the presence of Mg$\^$2+/ ions, suggesting that the metal ion stabilizes the stem-loop structure of the substrate RNA in solution. When point-mutated RNA substrates were used in the RNA cleavage assay and RNase T1 mapping study, the specific nucleotide sequences in the stem-loop region were not required for the accurate RNA cleavage by the viral capsid protein, but the formation of a stem-loop like structure in a region (nucleotides from 267 to 287) stabilized by Mg$\^$2+/ ions was critical for the accurate RNA cleavage. The RNase T1 mapping and EMSA studies revealed that the Ca$\^$2+/ and Mn$\^$2+/ ions, among the reagents tested, could change the mobility of the substrate RNA 5' 249-342 on a gel similarly to that of Mg$\^$2+/ ions, but only Ca$\^$2+/ ions identically showed the stabilizing effect of Mg$\^$2+/ ions on the stem-loop structure, suggesting that binding of the metal ions (Mg$\^$2+/ or Ca$\^$2+/) onto the RNA substrate in solution causes change and stabilization of the RNA stem-loop structure, and only the substrate RNA with a rigid stem-loop structure in the essential region can be accurately cleaved by the LRV1-4 viral capsid protein.

Molecular Cloning and High-Level Expression of Human Cytoplasmic Superoxide Dismutase Gene in Escherichia coli (사람의 세포질 Superoxide Dismutase 유전자의 클로닝과 대장균내에서의 대량발현에 관한 연구)

  • 이우길;김영호;양중익;노현모
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.91-97
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    • 1990
  • Complementary DNA (cDNA) coding for human cytoplasmic superoxide dismutase (SOD1) (superoxide: superoxide oxidoreductase E.C.1.15.1.1) was isolated from human liver cDNA library of $\lambda$gt11 by in situ plaque hybridization. The insery cDNA gas the 5' untranslational region (UTR) and 3'UTR of SOD1 gene. Polymerase Chain Reaction (PCR) method was used fro subcloning of SOD1 structural gene. Using synthetic sense strand primer (24mer) containing a start codon and antisense strand primer (24mer), SOD1 structural gene was selectively amplified. Amplified DNA was directly cloned into the HincII site of pUC19 plasmid. Insery cDNA was subcloned into M13 mp19 and sequenced by dideowy chain termination method with Sequenase. The nucleotide sequence of insert cDNA had an open reading frame (ORF) coding for 153 amino acid residues. The structural gene of cytoplasmic SOD was placed under the control of bacteriophage $\lambda P_{L}$ regulatory sequences, generating a highly efficient expression plasmid. The production of human SOD1 in E. coli cells was about 7% of total cellular proteins and recombinant human SOD1 possessed its own enzymatic acitivity.

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Porcine LMNA Is a Positional Candidate Gene Associated with Growth and Fat Deposition

  • Choi, Bong-Hwan;Lee, Jung-Sim;Lee, Seung-Hwan;Kim, Seung-Chang;Kim, Sang-Wook;Kim, Kwan-Suk;Lee, Jun-Heon;Seong, Hwan-Hoo;Kim, Tae-Hun
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.12
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    • pp.1649-1659
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    • 2012
  • Crosses between Korean and Landrace pigs have revealed a large quantitative trait loci (QTL) region for fat deposition in a region (89 cM) of porcine chromosome 4 (SSC4). To more finely map this QTL region and identify candidate genes for this trait, comparative mapping of pig and human chromosomes was performed in the present study. A region in the human genome that corresponds to the porcine QTL region was identified in HSA1q21. Furthermore, the LMNA gene, which is tightly associated with fat augmentation in humans, was localized to this region. Radiation hybrid (RH) mapping using a Sus scrofa RH panel localized LMNA to a region of 90.3 cM in the porcine genome, distinct from microsatellite marker S0214 (87.3 cM). Two-point analysis showed that LMNA was linked to S0214, SW1996, and S0073 on SSC4 with logarithm (base 10) of odds scores of 20.98, 17.78, and 16.73, respectively. To clone the porcine LMNA gene and to delineate the genomic structure and sequences, including the 3'untranslated region (UTR), rapid amplification of cDNA ends was performed. The coding sequence of porcine LMNA consisted of 1,719 bp, flanked by a 5'UTR and a 3'UTR. Two synonymous single nucleotide polymorphisms (SNPs) were identified in exons 3 and 7. Association tests showed that the SNP located in exon 3 (A193A) was significantly associated with weight at 30 wks (p<0.01) and crude fat content (p<0.05). This association suggests that SNPs located in LMNA could be used for marker-assisted selection in pigs.

Incidence of Aphid-Transmitted Strawberry Viruses in Korea and Phylogenetic Analysis of Korean Isolates of Strawberry Mottle Virus (진딧물 전반 딸기 바이러스 발생조사 및 딸기모틀바이러스의 계통분석)

  • Kwon, Sun-Jung;Yoon, Jung-Beom;Cho, In-Sook;Yoon, Ju-Yeon;Kwon, Tae-Ryong
    • Research in Plant Disease
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    • v.25 no.4
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    • pp.226-232
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    • 2019
  • Among more than 30 viruses infecting strawberry, aphid-transmitted viruses including Strawberry mild yellow edge virus (SMYEV), Strawberry mottle virus (SMoV), Strawberry crinkle virus (SCV), and Strawberry vein banding virus (SVBV) have been considered as the most economically important viruses of strawberry in the world. To determine the incidence of these four viruses in major Korean strawberry cultivars, field surveys were conducted in major production areas during 2018-2019. In our surveys, SMYEV and SMoV were detected with low infection rates of 0.7% and 1.3%, respectively, whereas SCV and SVBV were not detected. No obvious symptoms were observed in the strawberry plants infected by SMYEV or SMoV. Since no sequences of SMoV Korean isolates have been reported, we initially determined nucleotide sequence of the 3' untranslated region (UTR) of seven SMoV isolates obtained during the surveys. The 3' UTR sequences (782 nt) of seven Korean isolates were phylogenetically compared with those of the previously reported SMoV isolates. Phylogenetic analysis revealed that most Korean isolates are closely related to Canadian isolates and only little evolutionary differentiation occurred among the Koreans isolates. This might be due to the low incidence of SMoV in strawberry in Korea.

Effects of the cis-Acting Element in the 3' End of Porcine $\beta$-Casein Gene on the Expression in Mammary Epithelial Cells (돼지 $\beta$-Casein 유전자의 3' 말단 부위의 cis-Acting Element가 유선 상피 세포내의 발현에 미치는 영향)

  • Lee, Hwi-Cheul;Kim, Byoung-Ju;Byun, Sung-June;Lee, Seung-Hoon;Kim, Min-Ji;Chung, Hee Kyoung;Lee, Hyun-Gi;Jo, Su-Jin;Chang, Won-Kyong;Park, Jin-Ki;Lee, Poong-Yeon
    • Reproductive and Developmental Biology
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    • v.32 no.3
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    • pp.153-158
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    • 2008
  • Tissue-specific and temporal regulation of milk protein gene expression is advantageous when creating transgenic animal that produces foreign protein into milk. Gene expression, i.e. protein production, is regulated not only by promoter strength but also mRNA stability. Especially, poly A tail length by polyadenylation affects in vivo and in vitro mRNA stability and translation efficiency of the target gene. In the present study, nucleotide sequence of 3'-UTR was analyzed to evaluate the effects of mRNA stability on the target gene expression. Based on the poly A signal of 3' -untranslated region (UTR), nucleotide sequences of putative cytoplasmic polyadenylation elements (CPEs) and downstream elements (DSEs: U-rich, G-rich, GU-rich) were analyzed and used to construct 15 luciferase reporter vectors. Each vector was transfected to HC11 and porcine mammary gland cell (PMGC) and measured for dual luciferase expression levels after 48 hours of incubation. Luciferase expression was significantly higher in construct #6 (with CPE 2, 3 and DSE 1 of exon 9) and #11 (with CPE 2, 3 and DSE 1, 2 and 3 of exon 9) than construct #1 in the PMGC. These results suggest that expression of target genes in PMGC may be effectively expressed by using the construct #6 and #11 on production of transgenic pig.

Genetic Diversity of Sweet potato feathery mottle virus from Sweet Potatoes in Korea

  • Kwak, Hae-Ryun;Kim, Mi-Kyeong;Jung, Mi-Nam;Lee, Su-Heon;Park, Jin-Woo;Kim, Kook-Hyung;Ko, Sug-Ju;Choi, Hong-Soo
    • The Plant Pathology Journal
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    • v.23 no.1
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    • pp.13-21
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    • 2007
  • Sweet potato feathery mottle virus(SPFMV) is one of the most prevalent viruses infecting sweet potatoes and occurs widely in sweet potato cultivating areas in Korea. To assess their genetic variation, a total of 28 samples infected with SPFMV were subjected to restriction fragment length polymorphism(RFLP) analysis using DNAs amplified by RT-PCR with specific primer sets corresponding to the coat protein(CP) region of the virus. The similarity matrix by UPGMA procedure indicated that 28 samples infected with SPFMV were classified into three groups based on the number and size of DNA fragments by digestion of CP-encoding regions with 7 enzymes including SalI, AluI, EcoRI, HindIII, FokI, Sau3AI, and DraI bands. Four primer combinations out of 5 designed sets were able to differentiate SPFMV and sweet potato virus G infection, suggesting that these specific primers could be used to differentiate inter-groups of SPFMV. Sequence analysis of the CP genes of 17 SPFMV samples were 97-99% and 91-93% identical at the intra-group and inter-groups of SPFMV, respectively. The N-terminal region of the CP is highly variable and examination of the multiple alignments of amino acid sequences revealed two residues(residues 31 and 32) that were consistently different between SPFMV-O and SPFMV-RC.