• Title/Summary/Keyword: Putative promoter

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Isolation and Characterization of the mheA (Most Highly Expressed) Gene of Aspergillus oryzae

  • Liu, Pengcheng;Lim, Ji-Young;Kim, Hee-Seo;Kim, Jong-Hwa;Chae, Keon-Sang
    • Mycobiology
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    • v.40 no.3
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    • pp.208-209
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    • 2012
  • The amino acid sequence of the mheA gene of Aspergillus oryzae encodes a putative metallothionein-like protein 1. The size of the mheA transcript was 497 nt and the mheA promoter was induced by glucose, consistent with results of analysis by Northern hybridization and with the pdcA promoter, respectively.

Choristoneura fumiferana Granulovirus pk-1: A Baculoviral Protein Kinase

  • Giannopoulos, Paresa N.;Nassoury, Nasha;Lamontagne, Lucie;Guertin, Claude;Rashidan, Kianoush Khajeh
    • BMB Reports
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    • v.38 no.4
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    • pp.457-467
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    • 2005
  • Open reading frame (ORF) 3 on the Choristoneura fumiferana granulovirus (ChfuGV), located in the 11 kb fragment of the BamHI genomic bank encodes a predicted 32-kDa putative kinase protein. Bioinformatics analysis on the predicted amino acid sequence of ChfuGV PK-1 revealed the existence of 11 catalytic subdomains. Sequence analysis within the 5'-untranslated region (5'-UTR) of ChfuGV pk-1 indicates the presence of both putative early and late promoter motifs, indicating that pk-1 may be expressed throughout the infection cycle. Promoter sequence analysis reveals that pk-1 is deprived of a TATA box and appears instead to be regulated by other cis-acting transcriptional regulatory elements. Temporal transcription analysis by RT-PCR confirms the appearance of transcripts detected from 2 h p.i. until 72 h p.i. Northern blot hybridization characterizes pk-1 transcription as a 1.2 kb transcript. Homology comparisons reveal that ChfuGV PK-1 protein is most closely related to Phthorimaea operculalla granulovirus (PoGV) with 80% amino acid identity.

Analysis of Flavonoid 3',5'-Hydroxylase Gene in Transgenic Petunia (Petunia hybrida) Plants (형질 전환된 페튜니아 식물체에서의 Flavonoid 3',5' -Hydroxylase 유전자의 분석)

  • 김영희
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.323-327
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    • 1998
  • The flavonoid biosynthetic pathway has been studied as a genetic model system, particularly in Petunia hybrida. In order to study the flavonoid biosynthetic pathway, we constructed a fusion gene system between Cauliflower Mosaic Virus (CaMV) 35S promoter and eggplant flavonoid 3', 5'-hydroxylase in pBI 121 plasmid. An optimal condition for plant regeneration was observed when internode explants were cultured on MS medium supplemented with IAA 0.2 mg/L plus BA 3 mg/L. For plant transformation internode explants of Petunia hybrida were precultured on BM medium supplemented with IAA 0.2 mg/L plus BA 3 mg/L. Putative transgenic plants were selected on medium containing kanamycin 50 mg/L plus cefotaxim 300 mg/L. Putative selected transformants were confirmed by amplification of selectable marker gene (nptII) by polymerase chain reaction (PCR) and Southern hybridization of flavonoid 3',5'-hydroxylase gene.

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Functional Expression of SAV3818, a Putative TetR-Family Transcriptional Regulatory Gene from Streptomyces avermitilis, Stimulates Antibiotic Production in Streptomyces Species

  • Duong, Cae Thi Phung;Lee, Han-Na;Choi, Si-Sun;Lee, Sang-Yup;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.2
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    • pp.136-139
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    • 2009
  • Avermectin and its analogs are major commercial antiparasitic agents in the fields of animal health, agriculture, and human infections. Previously, comparative transcriptome analysis between the low-producer S. avermitilis ATCC31267 and the high-producer S. avermitilis ATCC31780 using a S. avermitilis whole genome chip revealed that 50 genes were overexpressed at least two-fold higher in S. avermitilis ATCC31780. To verify the biological significance of some of the transcriptomics-guided targets, five putative regulatory genes were individually cloned under the strong-and-constitutive promoter of the Streptomyces expression vector pSE34, followed by the transformation into the low-producer S. avermitilis ATCC31267. Among the putative genes tested, three regulatory genes including SAV213, SAV3818, and SAV4023 exhibited stimulatory effects on avermectin production in S. avermitilis ATCC31267. Moreover, overexpression of SAV3818 also stimulated actinorhodin production in both S. coelicolor M145 and S. lividans TK21, implying that the SAV3818, a putative TetR-family transcriptional regulator, could be a global upregulator acting in antibiotic production in Streptomyces species.

Expression of GFP Gene Driven by the Olive Flounder (Paralichthys olivaceus) hsc70 Promoter in Trangenic Medaka (Oryzias latipes) (넙치 (Paralichthys olivaceus) 열충격 유전자 hsp70 조절부위에 의한 형광단백질의 발현)

  • Lee, Jeong-Ho;Kim, Jong-Hyun;Noh, Jae Koo;Kim, Hyun Chul;Kim, Woo-Jin;Kim, Young-Ok;Kim, Kyung-Kil
    • Korean Journal of Ichthyology
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    • v.19 no.4
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    • pp.266-273
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    • 2007
  • Heat shock proteins (HSPs) are a family of highly conserved proteins playing an important role in the functioning of unstressed and stressed cells. The HSP70 family, the most widely studied of the hsps, is constitutively expressed (hsc70) in unstressed cells and is also induced in response to stressors (hsp70), especially those affecting the protein machinery. The HSP/HSC70 proteins act as molecular chaperones and are crucial for protein functioning, including folding, intracellular localization, regulation, secretion, and protein degradation. Here, we report the identification and characterization of the putative amino acid sequence deduced from one cDNA clone identified as heat shock protein 70. The alignment showed that the putative sequence is 100% identical to the heat shock protein 70 cognate (HSC 70) of olive flounder. The 5'-flanking region sequence (approximately 1 kb) ahead of the hsc70 gene was cloned by genome walking and a putative core promoter region and transcription elements were identified. We characterized the promoter of the olive flounder hsc70 gene by examining the ability of 5'-upstream fragments to drive expression of green fluorescent protein (GFP) in live embryos.

Analysis of Upstream Regulatory Region from Populus nigra × P. maximowiczii by Inverse PCR Technique (Inverse PCR 기법(技法)을 이용(利用)한 양황철 DNA의 Regulatory Region의 탐색(探索))

  • Son, Suk Gyu;Hyun, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.87 no.3
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    • pp.334-340
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    • 1998
  • This research was conducted to identify plant regulatory regions by gene tagging method. A promoterless GUS coding sequence was introduced to Populus nigra ${\times}$ P. maximowiczii via Agrobacterium strains(LBA4404/EHA101), and putative transgenic poplars were selected by culturing on medium containing G418($60mg/{\ell}$) and by GUS assay. Among them one positive plant was to amplify the native sequences flanking to the introduced GUS gene in plant genome by inverse PCR method and from this 730 by DNA product was obtained. After subcloning and sequencing, it has 88% homology to the Eucalyptus gunnii CAD(cinnamyl alcohol dehydrogenase) gene. The GUS gene fused with the putative promoter reinserted into poplar leaves by particle bombardment method to test the funtional promoter activity. Upon staining with X-gluc, many blue spots appeared on the leaf segments bombarded by the chimeric gene 2-3 days, thus the isolated DNA fragment contain some possible coding region as well as a putative regulatory sequences of poplar CAD gene.

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Expression of \beta-agarase Gene and Carabolite Repression in Escherichia coli by the Promoter of Alginate Lyase Gene Isolated from Marine Pseudomonas sp. (해양의 Pseudomonas sp. 로부터 분리한 alginate lyase 유전자의 promoter에 의한 대장균 내에서의 \beta-agarase 유전자의 발현과 catabolite repression의 변화)

  • 공인수;박제현;한정현;최윤혁;이종희;진철호;이정기
    • Microbiology and Biotechnology Letters
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    • v.29 no.2
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    • pp.72-77
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    • 2001
  • Expression of f3 ~agarase Gene and Catabolite Repression in Escherichia coli by the Promoter of Alginate Lyase Gene Isolated from Marine Pseudomonas sp. Jin, Cheal~Ho, J~Hyeon Park, Jeong-Hyun Han, YoonM Hyeok Chae, Jong~Hee Lee, Jung-Kee Lee!, and In-800 Kong*. Faculty of Food Science and Biotechnology, Pukyong National UniversitYt Pusan 608-737, Korea, llnBioNet Co. 1690-3 Taejon 306-230, Korea - Promoter is a key factor for expression of the recombinant protein. There are many promoters for overexpression of protein in various organisms. The aly promoter of Pseudomonas sp. W7 isolated from marine environment was known to be a constitutive expression promoter of the alginate lyase gene, and it's promoter activity is repressed by glucose in Escherichia coli. To investigate the catabolite repression of the aly promoter ~md association between the promoter mutants, f3 agarase gene, which was also cloned from Pseudomonas sp. W7 was connected to the aly promoter with the sequence the coding 46 N-terminal amino acids ofthe alginate lyase gene. The constructed plasmid was introduced into E. coli and the agarase activity was measured. Fourty six amino acids of the alginate lyase gene was serially deleted using peR to the direction of 5' upstream region and subcloned. The agarase was overexpressed by the aly promoter and the production of agarase was repressed by the addition of glucose into culture media. Fourty six amino acids of alginate lyase did not affect the production of agarase at all. The deletion of a putative stem-loop structure in the aly promoter induced the decrease of f3 -agarase productivity.

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Nucleotide Sequence Analyses of p10 Gene and its Promoter of Hyphantria cunea Nuclear Polyhedrosis Virus (Hyphantria cunea Nuclear Polyhedrosis Virus p10유전자와 프로모터의 염기서열 결정)

  • Park, Sun-A;Cha, Sung-Chul;Chang, Jae-Hyeok;Lee, Hyung-Hoan
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.131-137
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    • 1996
  • The sequences of p10 gene its promoter of Hyphantria cunea NPV were determined. According to the sequence analysis, the putative p10 gene ORF has 285 bp. The 5'-non-coding leader sequence of the p10 gene promoter contained the TATA box and the putative transcription initiation site TAAG motif. Poly (A) tail signals, AATAAA sequence was at site 65 base upstream from the 3' terminus. The deduced amino acid sequence of p10 protein was 95 with a predicted molecular weight of 10.26 kDa. In the p10 protein sequence, a hydrophobic region was present at the N-terminus of the protein, whereas the C-terminus was highly hydrophilic. The p10 protein of H. cunea NPV did not contain cysteine, histidine, trytophan, tryptophane, tyrosine, glutamine and asparagine residues.

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Transcriptional activation of an anti-oxidant mouse Pon2 gene by dexamethasone

  • Lim, Ji-Ae;Kim, Sang-Hoon
    • BMB Reports
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    • v.42 no.7
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    • pp.421-426
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    • 2009
  • Glucocorticoids regulate multiple physiological processes such as metabolic homeostasis and immune response. Mouse Pon2 (mPon2) acts as an antioxidant to reduce cellular oxidative stress in cells. In this present study, we investigated the transcriptional regulation of mPon2 by glucocorticoids. In the presence of glucocorticoid analogue dexamethasone, the expression of mPon2 mRNA in cells was increased, whereas the expression was inhibited by a transcription inhibitor actinomycin D. Glucocorticoid receptors bound to the putative glucocorticoid response elements located between -593 bp and -575 bp of the mPon2 promoter. Transcriptional activity was completely blocked when the putative element was mutated. Taken together, these results suggest that the expression of the mPon2 gene is directly regulated by glucocorticoid-glucocorticoid receptor complexes.

Comparison of enzymatic activities between the recombinant CHT1 proteins from the hard tick Haemaphysalis longicornis expressed in E. coli and baculovirus-mediated Sf 9 cells (E. coli와 baculovirus-mediated Sf 9 세포에서 발현된 진드기 H. longicornis의 CHT1 단백의 효소활성 비교)

  • You, Myung-Jo;Fujisaki, Kozo
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.139-144
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    • 2003
  • A chitinase cDNA named CHT1 was cloned from the hard tick, Haemaphysalis longicornis, and the enzymatic properties of its recombinant proteins were characterized. The CHT1 cDNA encodes 930 amino-acid (aa) residues including a 22 aa putative signal peptide, with the calculated molecular mass of the putative mature protein 104 kDa. The E coli-expressed rCHT1 exhibited weak chitinolytic activity against $4MU-(GlcNAc)_3$. The rCHT1 protein with higher activity was obtained using recombinant Autographa californica multiple nuclear polyhedrosis virus (AcMNPV), which expresses rCHT1 under polyhedrin promoter. These findings suggest that the rCHT1 expressed in baculovirus-mediated Sf 9 cells has a high activity than E coli-expressed rCHT1.