• Title/Summary/Keyword: 3 'UTR

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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.

Cloning and expression of Lipomyces starkeyi dextranase-encoding gene in yeasts

  • Kang, Hee-Kyoung;Park, Ji-Young;An, Joon-Seob;Kim, Seung-Heuk;Kim, Do-Man
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.402-406
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    • 2005
  • Lipomyces starkeyi produces a novel glucanhydrolase containing endo-dextranase and ${\alpha}-amylase$ activities. A cDNA from L. starkeyi encoding a dextranase was isolated and characterized. The 2,052 kb cDNA fragment (lsd1) carrying dextranase gene showed one open reading frame (ORF) composed of 1,824 bp flanked by a 41 bp 5'-UTR and a 184 bp 3'-UTR including a poly(A) tail of 27 bp. The ORF encodes for a 608 amino acid with a predicted molecular mass of 67.6 kDa. There was 77% deduced amino acid sequence identity between the LSD1 dextranase and the dextranase from Penicillium minioluteum. The primary structure of the dextranase from L. starkeyi has distant similarity with enzymes belonging to glycosyl hydrolase family 49. The lsd1 protein was expressed in the Saccharmyces cerevisiae under control of GAL1 promoter and active dextranase was produced.

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Genetic Characterization of Bovine Viral Diarrhea Virus from Korean Indigenous Calves in Gyeongbuk Province (경북지방에서 사육되고 있는 한우에서 소 바이러스성 설사 바이러스의 유전적 특징)

  • Song, Moo-Chan;Choi, Kyoung-Seong
    • Journal of Veterinary Clinics
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    • v.27 no.3
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    • pp.220-224
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    • 2010
  • Bovine viral diarrhea virus (BVDV) is an important worldwide disease in the livestock industry. To characterize BVDV circulating in Gyeongbuk province in the Republic of Korea which has the highest cattle population density, 365 rectal swabs from clinically BVDV Korean indigenous calves were collected. Fifty cases were identified as positive for BVDV. A phylogenetic analysis of 5'- untranslated regions (UTR) revealed that most of our cases belonged to BVDV-2a (n = 48), while only two cases were classified as BVDV-1a (n = 1) and 1b (n = 1), respectively. These results indicated that BVDV-2a is the most prevalent subgroup in Korean indigenous calves of Gyeongbuk province.

Identification of Novel Alternatively Spliced Transcripts of RBMS3 in Skeletal Muscle with Correlations to Insulin Action in vivo

  • Lee, Yong-Ho;Tokraks, Stephen;Nair, Saraswathy;Bogardus, Clifton;Permana, Paska A.
    • Biomedical Science Letters
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    • v.15 no.4
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    • pp.301-307
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    • 2009
  • Whole-body insulin resistance results largely from impaired insulin-stimulated glucose disposal in skeletal muscle. Our previous studies using differential display and quantitative real-time RT-PCR have shown that a novel cDNA band (DD23) had a higher level of expression in insulin resistant skeletal muscle and it was correlated with whole-body insulin action, independent of age, sex, and percent body fat. In this study, we cloned and characterized DD23. The DD23 sequence is part of the 3'UTR region of the RNA binding motif, single stranded interacting protein (RBMS3). We have cloned the full length cDNA for RBMS3 and identified two splice variants. These variants named DD23-L and DD23-S have 15 and 14 exons respectively and differ from RBMS3 in the 3'UTR significantly. Northern blot analyses showed that an ~8.8 kb mRNA transcript of DD23 was predominantly expressed in skeletal muscle and to a lesser extent in placenta, but not in heart, brain, lung, liver, or kidney, unlike RBMS3. Elevated expression levels of these novel alternatively spliced variants of RBMS3 in skeletal muscle may play a role in whole body insulin resistance.

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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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    • v.15 no.5
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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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Influence of Food Allowance on Nutrient Utilization of Oat (Avena sativa) Hay by Goats and Feeding Value of Leftovers from High Levels of Offer in Sheep

  • Dutta, N.;Sharma, K.;Hasan, Q.Z.;Pathak, N.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.5
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    • pp.723-727
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    • 1999
  • Two experiments were conducted to determine the effect of feeding levels on intake and nutrient utilization of oat (Avena sativa) hay (OH) by goats and potential feeding value of leftovers from high levels of offer in sheep. In experiment 1, the goats (15) were offered OH at three levels of feeding to give leftovers of about 20% (T-1), 35% (T-2) and 50% (T-3) of DM offered. A marked effect of refusal rate of OH on intake and digestibility of nutrients was evident. Allowing selective consumption at higher levels (T-2 and T-3), the intake of DCP and TDN from OH was found not only to meet the maintenance requirement but provided surplus nutrients for moderate production. In experiment 2, Sheep (3) were offered one of the three dietary treatments viz OH, refusals left uneaten by goats in T-2 and T-3 (OHR) and 3% urea treated oat hay refusals (UTR) in a $3{\times}3$ latin square design. Though the potential feeding value of leftovers (OHR and UTR) was lower than OH, the results confirm the possibility of their effective use. It was concluded that feeding above the conventional ad libitum level could be an alternative to improve the performance of ruminants fed feeds like oat hay. The potential impact of the strategy would, however, depend on the effective reutilization of leftovers for feeding other animals.

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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    • v.19 no.4
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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.

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.

Analysis of Single Nucleotide Polymorphisms of Leptin Gene in Hanwoo(Korean Cattle) (한우 Leptin 유전자의 단일 염기 다형성 분석)

  • Lee, J.-Min;Song, G.C.;Lee, J.Y.;Kim, Young-Bong
    • Journal of Animal Science and Technology
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    • v.49 no.3
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    • pp.295-302
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    • 2007
  • Leptin, the product of the obese(ob) gene, is an adipocyte-derived hormone for the regulation of whole- body energy storage and energy usage. It has been reported that the homozygous mutations in the gene for leptin(LEP) induce obesity and reduce energy expenditure. In cattle, LEP has significant roles directly or indirectly related with phenotypes such as body weight and fat deposits, therefore SNPs of LEP have been considered important genetic marker to estimate carcass fat content in cattle. In this study, SNPs were screened in LEP(2,222 bp) between intron 1 to 3'-UTR from 24 independent Hanwoo(Korean cattle) by PCR and DNA sequencing. Total 25 SNPs were found and two nonsynonymous SNPs including T1163A(V19E) and G3256A(G132D) were newly detected only from Hanwoo. Among 20 SNPs previously reported in cattle, 16 SNPs were found in Hanwoo; however, the frequencies of some SNPs were significantly different between Hanwoo and western cattle breeds. The other 4 SNPs were not detected from Hanwoo. These Hanwoo specific SNP patterns in LEP will be used in development of molecular marker and application to genetic improvement of Hanwoo.

miRNA-183 Suppresses Apoptosis and Promotes Proliferation in Esophageal Cancer by Targeting PDCD4

  • Yang, Miao;Liu, Ran;Li, Xiajun;Liao, Juan;Pu, Yuepu;Pan, Enchun;Yin, Lihong;Wang, Yi
    • Molecules and Cells
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    • v.37 no.12
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    • pp.873-880
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    • 2014
  • In our previous study, miRNA-183, a miRNA in the miR-96-182-183 cluster, was significantly over-expressed in esophageal squamous cell carcinoma (ESCC). In the present study, we explored the oncogenic roles of miR-183 in ESCC by gain and loss of function analysis in an esophageal cancer cell line (EC9706). Genome-wide mRNA micro-array was applied to determine the genes that were regulated directly or indirectly by miR-183. 3'UTR luciferase reporter assay, RT-PCR, and Western blot were conducted to verify the target gene of miR-183. Cell culture results showed that miR-183 inhibited apoptosis (p < 0.05), enhanced cell proliferation (p < 0.05), and accelerated G1/S transition (p < 0.05). Moreover, the inhibitory effect of miR-183 on apoptosis was rescued when miR-183 was suppressed via miR-183 inhibitor (p < 0.05). Western blot analysis showed that the expression of programmed cell death 4 (PDCD4), which was predicted as the target gene of miR-183 by microarray profiling and bioinformatics predictions, decreased when miR-183 was over-expressed. The 3'UTR luciferase reporter assay confirmed that miR-183 directly regulated PDCD4 by binding to sequences in the 3'UTR of PDCD4. Pearson correlation analysis further confirmed the significant negative correlation between miR-183 and PDCD4 in both cell lines and in ESCC patients. Our data suggest that miR-183 might play an oncogenic role in ESCC by regulating PDCD4 expression.