• Title/Summary/Keyword: Flanking region

Search Result 182, Processing Time 0.191 seconds

Regulatory Sequences in the 5' Flanking Region of Goat β-Casein Gene

  • Huang, Mu-Chiou;Chao, Jiunn-Shiuan
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.14 no.11
    • /
    • pp.1628-1633
    • /
    • 2001
  • A goat ${\beta}$-casein gene was cloned and sequenced. Our previous study had determined the nucleotide sequences of the 5' flanking region and the structural gene including all 9 exons. In the present study, investigations were done on the regulatory sequences in the 5' flanking region of the goat ${\beta}$-casein gene by aligning and comparing it with the same gene from other mammals. The results showed that -200/-1 bp of the 5' flanking sequences contained six conserved clusters, in which the sites of gene expression regulated by the transcription factor and hormone might exist. It showed that fourteen glucocorticoid receptor elements, two cAMP responsive elements, two SV40 virus enhancer core sequences, two OCT-1 binding elements and one CTF/NF-1 binding element were dispersed in the 5' flanking region of goat ${\beta}$-casein gene. Our findings are perhaps valuable for the elucidation of the molecular mechanisms that control the expression of the goat ${\beta}$-casein gene.

Importance of Nucleotides Adjacent to the Core Region of Diphtheria tox Promoter/Operator

  • Lee, John-Hwa
    • Journal of Microbiology and Biotechnology
    • /
    • v.12 no.4
    • /
    • pp.622-627
    • /
    • 2002
  • Diphtheria toxin repressor (DtxR) binds to approximately 30 to 35-bp regions containing an interrupted 9-bp inverted repeat within a 19-bp core sequence. The core sequence is fairly conserved and critical for DtxR binding. The flanking regions that are consisted of 5 to 8 more of nucleotides from the core are also required for DtxR binding. The nucleotides in both flanking regions are A-T rich. To examine whether the A-T nucleotides in both flanking regions from the core have significant roles for DtxR binding, a DNA fragment was constructed based on the diphtheria tox promoter/operator, and DNA fragments with substitution of A and T nucleotides In the flanking regions to G and C were also constructed. To assess the effect of these substitutions on binding of DtxR and repressibility by DtxR, $\beta$-galactosidase activity from lacZ fused to the region was assessed. Gel mobility shift of the region by purified DtxR was also examined. The DNA fragments containing the mutations in the flanking regions still exhibited repression and mobility shift with DtxR. The core segment with the mutation is still, therefore, recognized by DtxR. Nonetheless, the results from the assays indicated that the substitution significantly decreased repression of the operator by DtxR in vivo under high-iron condition and decreased binding of DtxR to the operator. These results suggest that A and T nucleotides fur both flanking regions are preferred for the binding of DtxR.

Association of a Single Nucleotide Polymorphism in the 5'-Flanking Region of Porcine HSP70.2 with Backfat Thickness in Duroc Breed

  • Chen, Ming-Yu;Huang, San-Yuan;Lin, En-Chung;Hseu, Tzong-Hsiung;Lee, Wen-Chuan
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.16 no.1
    • /
    • pp.100-103
    • /
    • 2003
  • Higher environmental temperature affects the economic performance of pigs. Heat shock protein 70 has been shown to play an important role in thermoresistance. The purpose of this study was to assess the effect of a single nucleotide polymorphism in the 5'-flanking region of porcine HSP70.2 on growth performance in Taiwanese Duroc. The genotype of this nt 393 polymorphic site could be verified by digestion with Bsa WI restriction enzyme of a PCR product. Pigs with TT and TC genotypes have thinner backfats than those with CC type (p<0.05). The result suggested that the polymorphic Bsa WI site in the 5'flanking region of porcine HSP70.2 may be used as a marker for the early selection of ultrasonic backfat thickness in Duroc pigs.

Isolation of CD4 Genomic Clones and Role of Its 5' Upstream Region in CD4 Expression

  • Youn, Hyun-Joo
    • Korean Journal of Microbiology
    • /
    • v.30 no.6
    • /
    • pp.488-494
    • /
    • 1992
  • Three clones containing mouse CD4 gene were prepared using AKR genomic cosmid library. The role of 6, 500 bp 5' flanking region of the first exon of the AKR CD4 gene in tissue or developmental stage specific expression of CD4 has been studied. The deletion constructs containing various amounts of CD4 5' flanking sequences were prepared, and they were transfected into the cell lines representing different cell types or developmental stages of CD4 expression. Study of the reporter gene expression revealed that at least 1, 700 bp of 5' flanking region did retain promoter activity for CD4 expression. This area did not seem to contain enhancer activity for a full expression of CD4. However, the putative promoter interacted with other tissue specific enhancer sequence and showed the tissue specificity of the enhancer element.

  • PDF

Molecular Analysis of AQP2 Promoter. I. cAMP-dependent Regulation of Mouse AQP2 Gene

  • Park, Mi-Young;Lee, Yong-Hwan;Bae, Hae-Rahn;Lee, Ryang-Hwa;Lee, Sang-Ho;Jung, Jin-Sup
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.3 no.2
    • /
    • pp.157-164
    • /
    • 1999
  • To determine molecular mechanisms of Aquaporin-CD (AQP2) gene regulation, the promoter region of the AQP2 gene was examined by transiently transfecting a promoter-luciferase reporter fusion gene into mouse renal collecting duct cell lines such as mIMCD-3, mIMCD-K2, and M-1 cells, and NIH3T3 mouse embryo fibroblast cells. PCR-Southern analysis reveals that mIMCD-3 and mIMCD-K2 cells express AQP2, but M-1 and NIH3T3 cells do not, and that the treatment with cpt-cAMP $(400\;{\mu}M)$) or forskolin/isobutylmethylxanthine (IBMX) increased the AQP2 expression in IMCD cells. In both IMCD and NIH3T3 cells, the constructs containing the promoter of AQP2 gene showed promoter activities, indicating lack of tissue-specific element in the 1.4 kb 5'-flanking region of the mouse AQP2 gene. Luciferase activity in the IMCD cells transfected with the construct containing 5-flanking region showed responsiveness to cpt-cAMP, indicating that the 1.4 kb 5'-flanking region contains the element necessary for the regulatory mechanism by cAMP. The promoter-luciferase constructs which do not have a cAMP-responsible element (CRE) still showed the cAMP responsiveness in IMCD cells, but not in NIH3T3 cells. Increase in medium osmolarity did not affect AQP2 promoter activity in mIMCD-K2 cells. These results demonstrate that AQP2 gene transcription is increased with cAMP treatment through multiple motifs including CRE in the 5'-flanking region of the gene in vitro, and the regulatory mechanism may be important for in vivo regulation of AQP2 expression.

  • PDF

Characterization of the 5-Flanking region upstream from the structural gene for Zymononas mobilis alcohol dehydrogenase

  • Yoon, Ki-Hong;Park, Seung-Hwan;Jung, Kyung-Hwa;Pack, M. Y.
    • Journal of Microbiology
    • /
    • v.33 no.2
    • /
    • pp.126-127
    • /
    • 1995
  • A Zymomonas mobilis DNA fragment consisting of 207 nucleotides, which corresponded to the 5'-flanking region of an adhB gene encoding alcohol dehydrogenase II, was fused to the structural gene coding for a Bacillus endo-.betha.--1, 4-glucanase. The Z. mobilis DNA framgment waw identified to promote 50-fold increase in the expression of endo-.betha.1. 4 glucanase gene in Escherichia coli.

  • PDF

Nucleotide Sequence of a Truncated Proteinase Inhibitor I Gene of Potato (감자에서 분리된 절단형 단백질분해효소 억제제 I 유전자의 염기서열)

  • 이종섭
    • Journal of Plant Biology
    • /
    • v.33 no.4
    • /
    • pp.303-307
    • /
    • 1990
  • A genomic clone carrying a proteinase inhibitor I sequence was isolated and characterized. The clone contained a 0.7 kb EcoRI fragment hybridized with tomato inhibitor I cDNA. The nucleotide sequence of the EcoRI fragment revealed presence of a truncated form of a proteinase inhibitor I gene of potato. The truncated gene contained the 5' flanking region and the first exon of a functional proteinase inhibitor I gene. Although the 5' flanking region contained the regulatory sequences TATAAA and CCACT, a deletion of 40 bp occurred between them.

  • PDF

Nucleotide Sequences of β-lactoglobulin Gene 5'Flanking Region in Korean Native Goat (한국재래산양 β-lactoglobulin 유전자 5'flanking 영역의 염기서열 분석)

  • Ryoo, Seung-Heui;Han, Sung-Wook;Seo, Kil-Woong;Sang, Byung-Chan
    • Korean Journal of Agricultural Science
    • /
    • v.28 no.2
    • /
    • pp.78-84
    • /
    • 2001
  • This study was analyzed by PCR technique with specific primer in order to investigate the characteristics of ${\beta}$-lactoglobulin (${\beta}$-LG) gene 5'flanking region in Korean native goat. This work confirmed amplified product of 1,077 bp fragments obtained from the amplification of ${\beta}$-LG promoter from genomic DNA using PCR in Korean native goat. The nucleotide sequence of ${\beta}$-LG gene 5'flanking region in Korean native goat as compared with that of sheep ${\beta}$-LG were different at 46 base of 897 nucleotides, and showed high homology as about 94.9% each breed. Especially we confirmed that the difference of nucleotide sequences between Korean native goat and sheep were consisted of $T{\rightarrow}C$ substitution and $C{\rightarrow}T$ substitution reversely. As a consequences, the sequences of ${\beta}$-LG gene 5'flanking region showed a high homology between Korean native goat and sheep. Furthermore we should be studied that relationships between the control of gene expression and nucleotide sequences of transcription factor in Korean native goat.

  • PDF

Expression of a Small Protein Encoded by the 3' Flanking Sequence of the Escherichia coli rnpB Gene

  • Kim, Yool;Han, Kook;Lee, Jung-Min;Kim, Kwang-Sun;Lee, Young-Hoon
    • Bulletin of the Korean Chemical Society
    • /
    • v.28 no.6
    • /
    • pp.1010-1014
    • /
    • 2007
  • M1 RNA is the catalytic component of RNase P, a tRNA-processing enzyme in Escherichia coli. M1 RNA is produced in the cell by transcription of the rnpB gene and subsequent processing at the 3' end. The 3' flanking region of rnpB contains repeated sets of overlapping sequences coding for small proteins. The issue of whether these proteins are expressed remains to be established. In this study, we showed the expression of a small protein encoded by the first repeat within the 3' flanking region of rnpB. Interestingly, protein expression was increased at lower temperatures. The termination efficiency of rnpB terminators was decreased at lower temperatures, suggesting that antitermination is responsible for enhanced protein expression. Moreover, the purified small protein contained M1 RNA, implying a role as a specific RNA-binding protein.

Characterization of the Nanog 5'-flanking Region in Bovine

  • Choi, Don-Ho;Kim, Duk-Jung;Song, Ki-Duk;Park, Hwan-Hee;Ko, Tae Hyun;Pyao, Yuliya;Chung, Ku-Min;Cha, Seok Ho;Sin, Young-Su;Kim, Nam-Hyung;Lee, Woon-Kyu
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.29 no.10
    • /
    • pp.1383-1391
    • /
    • 2016
  • Bovine embryonic stem cells have potential for use in research, such as transgenic cattle generation and the study of developmental gene regulation. The Nanog may play a critical role in maintenance of the undifferentiated state of embryonic stem cells in the bovine, as in murine and human. Nevertheless, efforts to study the bovine Nanog for pluripotency-maintaining factors have been insufficient. In this study, in order to understand the mechanisms of transcriptional regulation of the bovine Nanog, the 5'-flanking region of the Nanog was isolated from ear cells of Hanwoo. Results of transient transfection using a luciferase reporter gene under the control of serially deleted 5'-flanking sequences revealed that the -134 to -19 region contained the positive regulatory sequences for the transcription of the bovine Nanog. Results from mutagenesis studies demonstrated that the Sp1-binding site that is located in the proximal promoter region plays an important role in transcriptional activity of the bovine Nanog promoter. The electrophoretic mobility shift assay with the Sp1 specific antibody confirmed the specific binding of Sp1 transcription factor to this site. In addition, significant inhibition of Nanog promoter activity by the Sp1 mutant was observed in murine embryonic stem cells. Furthermore, chromatin-immunoprecipitation assay with the Sp1 specific antibody confirmed the specific binding of Sp1 transcription factor to this site. These results suggest that Sp1 is an essential regulatory factor for bovine Nanog transcriptional activity.