• Title/Summary/Keyword: 3'-untranslated Region

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Characterization of the Small Cryptic Plasmid, pGD2, of Klebsiellia sp. KCL-2.

  • Yoo, Ju-Soon;Kim, Hae-Sun;Chung, Soo-Yeol;Lee, Young-Choon;Cho, Young-Soo;Choi, Yong-Lark
    • BMB Reports
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    • 제34권6호
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    • pp.584-589
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    • 2001
  • One of the cryptic plasmids from the oil degrading bacterium Klebsiella sp. KCL-2, the small plasmid pGD2, has been identified and characterized. This plasmid has a size of 3.6 kb with unknown functions. We constructed the recombinant plasmid pMGD2. The nucleotide sequences of the plasmid were determined and two open reading frames were detected. ORF1 encodes a replication initiator protein (RepA), which has a high degree of homology with the protein of ColE2 plasmid. The product encoded by ORF2 showed a high similarity with the transposase protein of IS5. IS5 is 1195 by long and contains an inverted terminal repetition of 16 bp with one mismatch. Stem-loop structures in the 5'untranslated region of the repA suggest that a putative gene, incA, is located in a complementary strand to the leader region of the repA mRNA.

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한국인 신생아 황달에서 UGT1A1 유전자의 1828G>A 단일염기다형성에 관한 연구 (1828G>A polymorphism of the UDP-glucuronosyltransferase gene (UGT1A1) for neonatal hyperbilirubinemia in Koreans)

  • 김자영;김미연;김지숙;김은령;윤서현;이희제;정주호
    • Clinical and Experimental Pediatrics
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    • 제49권1호
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    • pp.34-39
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    • 2006
  • 목 적 : 신생아 황달은 중국, 일본, 한국인 등 동양인에서 서양인에 비해 많이 발생하여 유전적인 요인이 있을 것으로 생각되며 빌리루빈 대사의 주된 효소인 Uridine diphosphate glucuronosyl transferase 유전자의 다형성이 관여한다고 보고되고 있다. 저자들은 빌리루빈 대사의 핵심 효소인 UGT1A1 유전자의 다형성이 한국인 신생아 황달과 어떤 연관성이 있는지 알아 보고자 본 연구를 시행하였다. 방 법 : 혈중 빌리루빈 수치가 12 mg/dL 이상의 건강하고, 위험인자가 없는 만삭아 중 신생아 황달 환아 80명과, 대조군 164명으로부터 혈액을 0.5 cc를 채취하여 DNA을 분리하였다. UGT1A1 유전자를 PCR로 증폭하여, 염기서열 분석 방법을 통해 UGT1A1 유전자 untranslated region의 1828G>A(rs 10929303) 단일염기다형성을 확인하였다. 결 과 : UGT1A1 유전자 3' untranslated region의 guanine에서 adenine으로의 1828G>A 단일염기다형성이 신생아 고빌리루빈군 중 총 혈청 빌리루빈이 12 mg/dL 이상인 80명 중 67명(83.8%)이 GG 유전형을 보였고 10명(12.5%)이 GA 이종접합, 3명(3.7%)이 AA 유전형을 나타냈다. 대조군 164명에서는 135명(82.3%)이 GG 유전형을 보였고, GA 이종접합은 16명(9.8%), AA 유전형은 13명(7.9%)으로 나타났다. 변이형 대립유전자 빈도는 고빌리루빈혈증군에서 10.0%로 대조군 12.8%와 비슷하였다(P=0.3687). 결 론 : 한국인 신생아 황달에서 UGT1A1 유전자의 1828G>A 다형성을 확인하였으나, 이는 고빌리루빈혈증군에서 대조군에서와 비슷한 빈도로 관찰되어 한국인 신생아 황달의 발생과 연관이 없을 것으로 생각된다.

Inhibition of Cell Proliferation and Migration by miR-509-3p That Targets CDK2, Rac1, and PIK3C2A

  • Yoon, Sena;Han, Eunji;Choi, Young-Chul;Kee, Honghwan;Jeong, Yongsu;Yoon, Jaeseung;Baek, Kwanghee
    • Molecules and Cells
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    • 제37권4호
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    • pp.314-321
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    • 2014
  • CDK2 is a key regulator of cell cycle progression. In this study, we screened for miRNAs targeting CDK2 using a luciferase-3'-untranslated region reporter assay. Among 11 hit miRNAs, miR-509-3p reduced CDK2 protein levels and significantly inhibited cancer cell growth. Microarray, Western blotting, and luciferase reporter analyses revealed additional targets of miR-509-3p, including Rac1 and PIK3C2A. Overexpression of miR-509-3p induced G1 cell-cycle arrest and inhibited colony formation and migration. RNAi experiments indicated that the growth-inhibitory effects of miR-509-3p may occur through down-regulation of CDK2, Rac1, and PIK3C2A. Targeting of multiple growth regulatory genes by miR-509-3p may contribute to effective anti-cancer therapy.

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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    • 제25권12호
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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.

수온변화에 따른 붉바리(Epinephelus akaara)의 heat shock protein (Hsp) 70 mRNA 발현 (Molecular Cloning and Expression Analysis of Red-spotted Grouper, Epinephelus akaara Hsp70)

  • 민병화;허준욱;박형준
    • 생명과학회지
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    • 제28권6호
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    • pp.639-647
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    • 2018
  • 한국의 고급 양식대상 어종인 붉바리(Epinephelus akaara)로부터 새로운 heat shock protein (Hsp) 70을 동정하였다. 붉바리 Hsp70 (RgHsp70)의 cDNA는 RACE (rapid amplification of cDNA ends)법을 사용하였고, RgHsp70 cDNA의 전장은 2,152 bp이고, 5'-terminal untranslated region (UTR)은 105 bp, 3'-terminal UTR은 274 bp, 590개의 아미노산을 암호화하는 open reading frame (ORF)는 1,773 bp였으며, 분자무게(molecular weight)는 64.9 kDa 및 등전위값(isoelectric point, pI)은 5.2였다. 추정되는 아미노산 비교 및 계통발생학적 분석 결과, 다른 어종과 마찬가지로 Hsp70 고유의 signature를 포함하는 것을 비롯하여 높은 유사성을 나타내었으므로 RgHsp70이 Hsp70 family임을 확인할 수 있었다 RgHsp70 mRNA는 간과 두신 조직에서 높은 발현을 보였으며, 48시간 동안 수온별(21, 18, 15 및 $12^{\circ}C$) 노출 후 간 조직에서 대조구인 $21^{\circ}C$보다 $12^{\circ}C$에서 발현이 증가함을 확인하였다. 본 연구에서는, 수온이 하강함에 따라 RgHsp70 mRNA 발현에 주요한 영향을 미치는 것으로 보아, 수온변화에 따른 스트레스로 인해 발현의 변화를 나타내는 주요 스트레스성 단백질임을 확인할 수 있었다.

Prediction of Mammalian MicroRNA Targets - Comparative Genomics Approach with Longer 3' UTR Databases

  • Nam, Seungyoon;Kim, Young-Kook;Kim, Pora;Kim, V. Narry;Shin, Seokmin;Lee, Sanghyuk
    • Genomics & Informatics
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    • 제3권3호
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    • pp.53-62
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    • 2005
  • MicroRNAs play an important role in regulating gene expression, but their target identification is a difficult task due to their short length and imperfect complementarity. Burge and coworkers developed a program called TargetScan that allowed imperfect complementarity and established a procedure favoring targets with multiple binding sites conserved in multiple organisms. We improved their algorithm in two major aspects - (i) using well-defined UTR (untranslated region) database, (ii) examining the extent of conservation inside the 3' UTR specifically. Average length in our UTR database, based on the ECgene annotation, is more than twice longer than the Ensembl. Then, TargetScan was used to identify putative binding sites. The extent of conservation varies significantly inside the 3' UTR. We used the 'tight' tracks in the UCSC genome browser to select the conserved binding sites in multiple species. By combining the longer 3' UTR data, TargetScan, and tightly conserved blocks of genomic DNA, we identified 107 putative target genes with multiple binding sites conserved in multiple species, of which 85 putative targets are novel.

Post-transcriptional and post-translational regulation during mouse oocyte maturation

  • Kang, Min-Kook;Han, Seung-Jin
    • BMB Reports
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    • 제44권3호
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    • pp.147-157
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    • 2011
  • The meiotic process from the primordial stage to zygote in female germ cells is mainly adjusted by post-transcriptional regulation of pre-existing maternal mRNA and post-translational modification of proteins. Several key proteins such as the cell cycle regulator, Cdk1/cyclin B, are post-translationally modified for precise control of meiotic progression. The second messenger (cAMP), kinases (PKA, Akt, MAPK, Aurora A, CaMK II, etc), phosphatases (Cdc25, Cdc14), and other proteins (G-protein coupled receptor, phosphodiesterase) are directly or indirectly involved in this process. Many proteins, such as CPEB, maskin, eIF4E, eIF4G, 4E-BP, and 4E-T, post-transcriptionally regulate mRNA via binding to the cap structure at the 5' end of mRNA or its 3' untranslated region (UTR) to generate a closed-loop structure. The 3' UTR of the transcript is also implicated in post-transcriptional regulation through an association with proteins such as CPEB, CPSF, GLD-2, PARN, and Dazl to modulate poly(A) tail length. RNA interfering is a new regulatory mechanism of the amount of mRNA in the mouse oocyte. This review summarizes information about post-transcriptional and post-translational regulation during mouse oocyte meiotic maturation.

Complete Sequence Analysis of a Korean Isolate of Chinese Yam Necrotic Mosaic Virus and Generation of the Virus Specific Primers for Molecular Detection

  • Kwon, Sun-Jung;Cho, In-Sook;Choi, Seung-Kook;Yoon, Ju-Yeon;Choi, Gug-Seoun
    • 식물병연구
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    • 제22권3호
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    • pp.194-197
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    • 2016
  • Chinese yam necrotic mosaic virus (CYNMV) is one of the most widespread viruses in Chinese yam (Dioscorea opposita Thunb.) and causes serious yield losses. Currently, genetic information of CYNMV is very restricted and complete genome sequences of only two isolates (one from Japan and another from China) have been reported. In this study, we determined complete genome sequence of the CYNMV isolate AD collected from Andong, Korea. Genetic analysis of the polyprotein amino acid sequence revealed that the Korean isolate AD has high similarity with the Japanese isolate PES3 (97%) but relatively low similarity with the Chinese isolate FX1 (78%). Phylogenetic analysis using the CYNMV 3' proximal nucleotide sequences harboring the coat protein and 3' untranslated region further supported genetic relationship among the CYNMV isolates. Based on comparative analysis of the CYNMV genome sequences determined in this study and other previous studies, we generated molecular detection primers that are highly specific and efficient for CYNMV diagnosis.

Molecular Cloning of the 3'-Terminal Region of Garlic Potyviruses and Immunological Detection of Their Coat Proteins

  • Song, Sang-Ik;Song, Jong-Tae;Chang, Moo-Ung;Lee, Jong-Seob;Park, Yang-Do
    • The Plant Pathology Journal
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    • 제15권5호
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    • pp.270-279
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    • 1999
  • cDNAs complementary to the 3'-terminal regions of two potyvirus genomes were cloned and sequenced. The clone G7 contains one open reading frame (ORF) of 1,338 nucleotides and a 3' untranslated region (3'-UTR) of 403 nucleotides at the 3'-end excluding the 3'end poly(A) tail. The putative viral coat protein (CP) shows 55%-92% amino acid sequence homology to those of Allium potyviruses. The genome size of the virus was analyzed to be about 9.0 kb by Northern blot analysis. Five cDNA clones were screened out using GPV2 oligonucleotide as a probe. One of these clones, DEA72, which has a longest cDNA insert, contains one ORF of 1,459 nucleotides and a 3'-UTR of 590 nucleotides at the 3'-end excluding the 3'-end poly(A) tail. The putative viral CP shows 57%-88% amino acid sequence homologies to those of Allium potyviruses. The genome size of the virus was analyzed to be about 9.6 kb by Northern blot analysis. The results of immunoblot and Northern blot analyses suggest that almost all of the tested garlic plants showing mosaic or streak symptoms are infected with DEA72-potyvirus in variable degrees but rarely infected with G7-potyvirus in variable degrees but rarely infected with DEA72-potyvirus in variable degrees but rarely infected with G7-potyvirus. Immunoelectron microscopy using anti-DEA72 CP antibody shows that this potyvirus is about 750 nm long and flexuous rod shaped.

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Genetic analysis of the postsynaptic transmembrane X-linked neuroligin 3 gene in autism

  • Hegde, Rajat;Hegde, Smita;Kulkarni, Suyamindra S.;Pandurangi, Aditya;Gai, Pramod B.;Das, Kusal K.
    • Genomics & Informatics
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    • 제19권4호
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    • pp.44.1-44.9
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    • 2021
  • Autism is a complex neurodevelopmental disorder, the prevalence of which has increased drastically in India in recent years. Neuroligin is a type I transmembrane protein that plays a crucial role in synaptogenesis. Alterations in synaptic genes are most commonly implicated in autism and other cognitive disorders. The present study investigated the neuroligin 3 gene in the Indian autistic population by sequencing and in silico pathogenicity prediction of molecular changes. In total, 108 clinically described individuals with autism were included from the North Karnataka region of India, along with 150 age-, sex-, and ethnicity-matched healthy controls. Genomic DNA was extracted from peripheral blood, and exonic regions were sequenced. The functional and structural effects of variants of the neuroligin 3 protein were predicted. One coding sequence variant (a missense variant) and four non-coding variants (two 5'-untranslated region [UTR] variants and two 3'-UTR variants) were recorded. The novel missense variant was found in 25% of the autistic population. The C/C genotype of c.551T>C was significantly more common in autistic children than in controls (p = 0.001), and a significantly increased risk of autism (24.7-fold) was associated with this genotype (p = 0.001). The missense variant showed pathogenic effects and high evolutionary conservation over the functions of the neuroligin 3 protein. In the present study, we reported a novel missense variant, V184A, which causes abnormal neuroligin 3 and was found with high frequency in the Indian autistic population. Therefore, neuroligin is a candidate gene for future molecular investigations and functional analysis in the Indian autistic population.