• Title/Summary/Keyword: 5' UTR

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Analysis of Trans-splicing Transcripts in Embryonic Stem Cell (배아줄기세포에서 트랜스 스플라이싱 전사체의 분석)

  • Ha, Hong-Seok;Huh, Jae-Won;Kim, Dae-Soo;Park, Sang-Je;Bae, Jin-Han;Ahn, Kung;Yun, Se-Eun;Kim, Heui-Soo
    • Journal of Life Science
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    • v.19 no.4
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    • pp.549-552
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    • 2009
  • Genetic mutations by gene fusion result from chromosomal rearrangement, trans-splicing, and intergenic splicing. Trans-splicing is a phenomenon in which two pre-mRNAs grow together into one. We analyzed the trans-splicing products in embryonic stem cells. By using bioinformatic tools, 70 trans-splicing transcripts were identified. They are classified into 6 types according to fusion pattern: 5'UTR-5'UTR, 5'UTR-3'UTR, 3'UTR-3'UTR, 5'UTR-CDS, 3'UTR-CDS, CDS-CDS. The fusion products are more abundant in CDS regions than in UTR regions, which contain multiple intron numbers. Chromosome analysis showing gene fusion via trans-splicing indicated that chromosomes 17 and 19 were activated. These data are of great use for further studies in relation to fusion genes and human diseases.

Toxicity of Tomato Spotted Wilt Virus Glycoprotein Signal Peptide and Promoter Activity of th 5' UTR

  • Park, Tae-Jin;Kim, Sun-Chang;Thomas L. German
    • The Plant Pathology Journal
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    • v.15 no.6
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    • pp.313-318
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    • 1999
  • Cloning of the 5'untranslated region (5' UTR) and Nterminus of the glycoprotein precursor (G2G1) open reading frame of tomato spotted wilt virus has been problematic, possibly because of the toxicity of a signal peptide at the beginning of th G2G1 protein precursor. The toxicity of the signal peptide to bacterial growth and the reason for the expression of the peptide gene in Escherichia coli were investigated by cloning the 5' UTR and the signal peptide sequence separately. Cells transformed with the plasmid containing both the first 30 amino acids of the glycoprotein and the 5' UTR showed a severe growth inhibition whereas transformants harboring either the plasmid with the signal sequence or the 5'UTR alone did not show any ingibition. An E. coli promoter-like sequence was found in the 5'UTR and tis promoter acivity was confirmed with a promoter-less GUS gene cloned downstream of the 5'UTR. In the cloning of the Tomato spotted wilt virus (TSWV) glycoprotein G2G1 open reading frame all the recovered plasmids contained stop codons in the signal sequence region. However, clones containing no stop codon were recovered when the signal sequence and the 5'UTR were cloned separately.

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Polymorphisms in the Thymidylate Synthase Gene and Risk of Colorectal Cancer

  • Gao, Chang-Ming;Ding, Jian-Hua;Li, Su-Ping;Liu, Yan-Ting;Cao, Hai-Xia;Wu, Jian-Zhong;Tajima, Kazuo
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.4087-4091
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    • 2012
  • To evaluate the relationship between polymorphisms (28 bp repeated sequences in 5'-UTR and 6-bp ins/del in 3'-UTR) in then thymidylate synthetase gene (TS) and risk of colorectal, colon and rectal cancers, we conducted a case-control study with 315 cases of colorectal cancer and 439 population-based controls in Jiangsu province, China. TS genotypes were identified using PCR.RFLP (restriction fragment length polymorphism) methods. Odds ratios (ORs) were estimated with an unconditional logistic regression model. We found that the distributions of 5'-UTR genotypes in TS were significantly different between controls and male colon cases (${\chi}^2$=8.25, P = 0.016). Compared with 3R/3R genotype, individuals with the 2R allele were at an increased risk of colon cancer (age-, BMI-, smoking- and alcohol drinking-adjusted OR=1.98, 95%CI: 1.11-3.53) among men. In ccontrast, the 6-bp ins/del polymorphism at the TS 3'- UTR did not influence risk of the colorectal, colon and rectal cancers. When combined genotypes for both TS 5'-UTR and 3'-UTR polymorphisms were evaluated, individuals with the 5'-UTR 2R allele had a OR of 3.61 (95%CI: 1.38-9.49) for colon cancer among men with the 3'-UTR .6bp/-6bp genotype. These results show that the polymorphism of the 28 bp repeated sequences in TS 5'-UTR could influence susceptibility to colon cancer and that there was a coordinated effect between TS 3'-UTR and 5'-UTR polymorphisms in increasing risk of colon cancer among Chinese men.

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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Isolation of an actin promoter for strong expression of transgenes in the orchid genus Dendrobium

  • Koo, Ja Choon
    • Journal of Plant Biotechnology
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    • v.40 no.1
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    • pp.27-36
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    • 2013
  • We isolated and functionally characterized a Dendrobium Actin1 (DmACT1) promoter that drives strong gene expression in the orchid genus Dendrobium. A genomic fragment containing the region 3227 bp upstream of the coding region of DmACT1 was obtained by inverse PCR. Detailed comparison of the full-length cDNA and genomic sequences revealed that DmACT1 has a 1374 bp first intron in the 5' UTR. However, the 5' flanking sequences upstream of the coding region showed no obvious sequence similarities compared to those of known promoters, including plant actin promoters. Serial deletion constructs of the 5' flanking region from the translation initiation codon were fused to the coding sequence of a GUS/luciferase fusion reporter to identify the regulatory elements necessary for promoter activity. Transient assays in the flowers of Dendrobium revealed that the 5' UTR-intron greatly enhanced promoter activity. Moreover, the DmACT1 promoter with its 5' UTR-intron yielded approximately 10-fold higher reporter activity than the rice Act1 promoter-intron. Our data suggest that the DmACT1 promoter with its 5' UTR-intron is a useful tool for strong expression of transgenes in Dendrobium orchids.

Secondary structure of the Irf7 5'-UTR, analyzed using SHAPE (selective 2'-hydroxyl acylation analyzed by primer extension)

  • Kim, Yun-Mi;Choi, Won-Young;Oh, Chang-Mok;Han, Gyoon-Hee;Kim, Young-Joon
    • BMB Reports
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    • v.47 no.10
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    • pp.558-562
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    • 2014
  • OASL1 is a member of the 2'-5'-oligoadenylate synthetase (OAS) family and promotes viral clearance by activating RNase L. OASL1 interacts with the 5'-untranslated region (UTR) of interferon regulatory factor 7 (Irf7) and inhibits its translation. To identify the secondary structure required for OASL1 binding, we examined the 5'-UTR of the Irf7 transcript using "selective 2'-hydroxyl acylation analyzed by primer extension" (SHAPE). SHAPE takes advantage of the selective acylation of residues in single-stranded regions by 1-methyl-7-nitroisatoic anhydride (1M7). We found five major acylation sites located in, or next to, predicted single-stranded regions of the Irf7 5'-UTR. These results demonstrate the involvement of the stem structure of the Irf7 5'-UTR in the regulation of Irf7 translation, mediated by OASL1.

Germline Variations of Apurinic/Apyrimidinic Endonuclease 1 (APEX1) Detected in Female Breast Cancer Patients

  • Ali, Kashif;Mahjabeen, Ishrat;Sabir, Maimoona;Baig, Ruqia Mehmood;Zafeer, Maryam;Faheem, Muhammad;Kayani, Mahmood Akhtar
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7589-7595
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    • 2014
  • Apurinic/apyrimidinic endonuclease 1 (APEX1) is a multifunctional protein which plays a central role in the BER pathway. APEX1 gene being highly polymorphic in cancer patients and has been indicated to have a contributive role in Apurinic/apyrimidinic (AP) site accumulation in DNA and consequently an increased risk of cancer development. In this case-control study, all exons of the APEX1 gene and its exon/intron boundaries were amplified in 530 breast cancer patients and 395 matched healthy controls and then analyzed by single-stranded conformational polymorphism followed by sequencing. Sequence analysis revealed fourteen heterozygous mutations, seven 5'UTR, one 3'UTR, two intronic and four missense. Among identified mutations one 5'UTR (rs41561214), one 3'UTR (rs17112002) and one missense mutation (Ser129Arg, Mahjabeen et al., 2013) had already been reported while the remaining eleven mutations. Six novel mutations (g.20923366T>G, g.20923435G>A, g.20923462G>A, g.20923516G>A, 20923539G>A, g.20923529C>T) were observed in 5'UTR region, two (g.20923585T>G, g.20923589T>G) in intron1 and three missense (Glu101Lys, Ala121Pro, Ser123Trp) in exon 4. Frequencues of 5'UTR mutations; g.20923366T>G, g.20923435G>A and 3'UTR (rs17112002) were calculated as 0.13, 0.1 and 0.1 respectively. Whereas, the frequency of missense mutations Glu101Lys, Ser123Trp and Ser129Arg was calculated as 0.05. A significant association was observed between APEX1 mutations and increased breast cancer by ~9 fold (OR=8.68, 95%CI=2.64 to 28.5) with g.20923435G>A (5'UTR), ~13 fold (OR= 12.6, 95%CI=3.01 to 53.0) with g.20923539G>A (5'UTR) and~5 fold increase with three missense mutations [Glu101Lys (OR=4.82, 95%CI=1.97 to 11.80), Ser123Trp (OR=4.62, 95%CI=1.7 to 12.19), Ser129Arg (OR=4.86, 95%CI=1.43 to 16.53)]. The incidence of observed mutations was found higher in patients with family history and with early menopause. In conclusion, our study demonstrates a significant association between germ line APEX1 mutations and breast cancer patients in the Pakistani population.

The Effect of Replacing Grass with Urea Treated Fresh Rice Straw in Dairy Cow Diet

  • Van Man, Ngo;Wiktorsson, Hans
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.8
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    • pp.1090-1097
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    • 2001
  • Nine tons of fresh rice straw from early-maturing varieties was ensiled with 50 g urea $kg^{-1}$ DM straw in plastic bags immediately after threshing. Five months after storage, preserved straw was used to partially replace elephant grass (Pennisetum purpureum, Schumach) for lactating cows. Eight crossbred Holstein lactating cows (75% of Holstein blood) in their second to fourth lactation and in mid-lactation were arranged in a balanced design with two squares consisting of $4\;periods{\times}4\;treatments$ (100% grass ad lib. as a control; 75% grass+urea treated fresh rice straw (UTrFRS) ad lib.; 50% grass+UTrFRS ad lib.; 25% grass+UTrFRS ad libitum) in one square. A concentrate supplement was given at a rate of 400 g per day per kg of milk produced. Samples of fresh straw taken in the field and UTrFRS and elephant grass taken at feeding time were evaluated in a degradation trial with 3 fistulated heifers (undefined blood ratio of crossbred of Sindhi and local yellow cattle). Straw preserved for 5-9 months was in nearly all cases of good quality. Crude protein (CP) content was increased 2.1 fold and 48 h dry matter loss (DML) was 20% higher compared to dry straw. Elephant grass cultivated intensively was low in DM content and 10% higher in 48 h DML compared to UTrFRS. Dry matter intake (DMI) was higher for the mixture of UtrFRS and Elephant grass, and highest when one-third of the roughage was UTrFRS. Higher DMI of mixed roughage diets was probably due to the low DM content of elephant grass in the sole grass roughage diet. Increasing substitution of elephant grass with UTrFRS up to 75% of the roughage component increased milk fat content and had no effect on milk yield and other milk composition parameters. Feeding UTrFRS, partially replacing elephant grass in the diets of lactating cows in the dry season can reduce the cost of roughage.

2R of Thymidylate Synthase 5'-untranslated Enhanced Region Contributes to Gastric Cancer Risk: a Meta-analysis

  • Yang, Zhen;Liu, Hong-Xiang;Zhang, Xie-Fu
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.1923-1927
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    • 2012
  • Background: Studies investigating the association between 2R/3R polymorphisms in the thymidylate synthase 5'-untranslated enhanced region (TYMS 5'-UTR) and gastric cancer risk have generated conflicting results. Thus, a meta-analysis was performed to summarize the data on any association. Methods: Pubmed, Embase, and CNKI databases were searched for all available studies. The strength of association between TYMS 5'-UTR 2R/3R polymorphism and gastric cancer risk was estimated by odds ratios (ORs) with 95% confidence intervals (CIs). Results: Six individual case-control studies with a total of 1, 472 cases and 1, 895 controls were included into this meta-analysis. Analyses of total six relevant studies showed that there was no obvious association between the TYMS 5'-UTR 2R/3R polymorphism and gastric cancer risk. Subgroup analyses based on ethnicity showed 2R of TYMS 5'-UTR 2R/3R contributes to gastric cancer risk in the Asian population ($OR_{Homozygote\;model}$ = 1.71, 95%CI 1.19-2.46, P = 0.004; $OR_{Recessive\;genetic\;model}$ = 1.70, 95%CI 1.18-2.43, P = 0.004). However, the association in Caucasian populations was uncertain due to the limited studies. Conclusions: Our meta-analysis suggests that 2R of TYMS 5'-UTR 2R/3R contributes to gastric cancer risk in the Asian population, while this association in Caucasians populations needs further study.

The relationship between the variants in the 5'-untranslated regions of equine chorionic gonadotropin genes and serum equine chorionic gonadotropin levels

  • Liu, ShuQin;Lian, Song;Yang, YunZhou;Fu, ChunZheng;Ma, HongYing;Xiong, ZhiYao;Ling, Yao;Zhao, ChunJiang
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.12
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    • pp.1679-1683
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    • 2017
  • Objective: An experiment was conducted to study the association between the single nucleotide polymorphisms (SNPs) in 5'-untranslated regions (5'-UTR) of equine chorionic gonadotropin (eCG) genes and the serum eCG levels. Methods: SNPs in 5'-UTR of eCG genes were screened across 10 horse breeds, including 7 Chinese indigenous breeds and 3 imported breeds using iPLEX chemistry, and the association between the serum eCG levels of 174 pregnant Da'an mares and their serum eCG levels (determined with ELISA) was analyzed. Results: Four SNPs were identified in the 5'-UTR of the $eCG{\alpha}$ gene, and one of them was unique in the indigenous breeds. There were 2 SNPs detected at the 5' end of the $eCG{\beta}$ subunit gene, and one of them was only found in the Chinese breeds. The SNP g.39948246T>C at the 5'-UTR of $eCG{\alpha}$ was associated significantly with eCG levels of 75-day pregnant mare serum (p<0.05) in Da'an mares. Prediction analysis on binding sites of transcription factors showed that the g.39948246T>C mutation causes appearance of the specific binding site of hepatocyte nuclear factor 3 forkhead homolog 2 (HFH-2), which is a transcriptional repressor belonging to the forkhead protein family of transcription factors. Conclusion: The SNP g.39948246T>C at the 5'-UTR of $eCG{\alpha}$ is associated with eCG levels of 75-day pregnant mare serum (p<0.05).