• Title/Summary/Keyword: open library

Search Result 805, Processing Time 0.024 seconds

Cloning and Characterization of a Novel Mannanase from Paenibacillus sp. BME-14

  • Fu, Xiaoyu;Huang, Xiaoluo;Liu, Pengfu;Lin, Ling;Wu, Gaobing;Li, Chanjuan;Feng, Chunfang;Hong, Yuzhi
    • Journal of Microbiology and Biotechnology
    • /
    • v.20 no.3
    • /
    • pp.518-524
    • /
    • 2010
  • A mannanase gene (man26B) was obtained from a sea bacterium, Paenibacillus sp. BME-14, through the constructed genomic library and inverse PCR. The gene of man26B had an open reading frame of 1,428 bp that encoded a peptide of 475- amino acid residues with a calculated molecular mass of 53 kDa. Man26B possessed two domains, a carbohydrate binding module (CBM) belonging to family 6 and a family 26 catalytic domain (CD) of glycosyl hydrolases, which showed the highest homology to Cel44C of P. polymyxa (60% identity). The optimum pH and temperature for enzymatic activity of Man26B were 4.5 and $60^{\circ}C$, respectively. The activity of Man26B was not affected by $Mg^{2+}$ and $Co^{2+}$, but was inhibited by $Hg^{2+},\;Ca^{2+},\;Cu^{2+},\;Mn^{2+},\;K^+,\;Na^+$, and $\beta$-mercaptoethanol, and slightly enhanced by $Pb^{2+}$ and $Zn^{2+}$. EDTA did not affect the activity of Man26B, which indicates that it does not require divalent ions to function. Man26B showed a high specific activity for LBG and konjac glucomannan, with $K_m,\;V_{max}$, and $k_{cat}$ values of 3.80 mg/ml, 91.70 ${\mu}mol$/min/mg protein, and 77.08/s, respectively, being observed when LBG was the substrate. Furthermore, deletion of the CBM6 domain increased the enzyme stability while enabling it to retain 80% and 60% of its initial activity after treatment at $80^{\circ}C$ and $90^{\circ}C$ for 30 min, respectively. This finding will be useful in industrial applications of Man26B, because of the harsh circumstances associated with such processes.

Long-term Monitoring Data for Growth and Yield of Local Rice Varieties in South Korea (국내 벼 지역별 주요 품종에 대한 장기 모니터링 자료의 구성형태)

  • Kim, Junhwan;Sang, Wangyu;Shin, Pyeong;Baek, Jaekyeong;Kwon, Dongwon;Lee, Yunho;Cho, Jung-Il;Seo, Myungchul
    • Korean Journal of Agricultural and Forest Meteorology
    • /
    • v.22 no.3
    • /
    • pp.176-182
    • /
    • 2020
  • National Institute of crop Science of the Rural Development Administration (RDA) has conducted long-term monitoring studies to find out the relationship between crop yield and climatic factors for major food crops including rice. Rice growth and y ield have been monitored in 17 regions where the branches of the National Institute of Crop science and the Provincial Agricultural Research and Extension Service locate. The data obtained from monitoring studies for rice growth and yield include the observation of vegetative growth status and yield components, which include leaf number, biomass and the weight of 1000 grains. These data have been collected from rice fields where standard management procedures have been applied. The observation data for crop growth and yield monitoring studies from 1999 to 2019 are open to public through agricultural science library operated by RDA.

Gene Cloning, Nucleotide Sequence and Efficent Expression of Peptidyl proryl cis-trans Isomerase from Bacillus stearothermophilus (Bacillus stearothermophilus의 Peptidyl Prolyl cis-trans Isomerase 유전자 분리 염기배열 및 발현)

  • 김동주
    • The Korean Journal of Food And Nutrition
    • /
    • v.9 no.4
    • /
    • pp.452-458
    • /
    • 1996
  • A PPIase gene of Bacillus stearothermophilus was screened from a genomic library by plaque hybridization using the A-1 primer as a probe. A PPIase positive plaque contained a 3.0kb insert of the chromosomal DNA. A 3.0kb fragment was subcloned into pUC18, resulting pPI1-40. A DNA fragment encoding the N-terminal portion of the PPIase in pPi-40 was amplified by polymerase chain reaction(PCR) method using the A-1 and B-2 primers. The amplified fragment was cloned into the Sma I site of pUC18 and recombinant plasmid was designated as pSN-18. The nucleotide sequence of 167bp fragment was determined. The deduced amino acid sequence of PPIase was completely matched with the determined N-terminal amino acid sequence of PPIase B. stearothermophilus. The translated protein sequence of PPIase B. stearothermophilus was compared with sequence from periplasmic PPIase from Escherichina coil ; homogies of 16 and 58%, respectively, were found. The clond PPIase gene was over-expressed in E. coil cell using pUC19 as an expression vector. The enzyme was partially purified by heat treatment and colum chromatochraphy on DEAE-Sepharose CL-6B. The molecular weight of the enzyme was dermined to be about 18.0 kDal by SDS-PAGE.

  • PDF

Expression and Cloning of the pmmC Gene Encoding Phosphomannomutase in Sphingomonas chungbukensis DJ77 (Sphingomonas chungbukensis DJ77 균주에서 Phosphomannomutase를 암호화하는 pmmC 유전자의 클로닝과 발현)

  • Kim Mi-Hye;Choi Jung-Do;Shin Malshick;Kim Young-Chang
    • Microbiology and Biotechnology Letters
    • /
    • v.33 no.2
    • /
    • pp.84-89
    • /
    • 2005
  • Phosphomannomutase (PMM) is a key enzyme in prokaryotes and eukaryotes, which catalyzes the conversion of ${\alpha}$-D-mannose 6-phosphate to ${\alpha}$-D-mannose 1-phosphate. The latter is the substrate for the synthesis of GDP-mannose, which serves as the mannosyl donor for many metabolic pathways in the cells. We report here on the isolation of a gene from a genomic library of Sphingomonas chungbukensis DJ77, the pmmC gene encoding phosphomannomutase. The gene was cloned into E. coli expression vector, and the sequence was analyzed. The ribosomal binding site GGAAG lays 5 bp upstream of the ORF of 750 bp, which is initiated by ATG codon and terminated by TAG. The predicted sequence of the enzyme consists of 249 amino acids with a molecular mass of 27.4 kDa and showed $86.9\%$ similarity to that of eukaryotic phosphomannomutase after bioinformatical analyses with the conserved domain search of NCBI. The purified gene product revealed the activity of phosphomannomutase. In conclusion, we confirmed that pmmC gene encodes phosphomannomutase actually.

Functional Genomics for Mass Analysis of Useful Genes in Panax ginseng C.A. Meyer (인삼의 유용유전자원 확보를 위한 기능 유전체연구)

  • Yang, Deok-Chun
    • Proceedings of the Ginseng society Conference
    • /
    • 2004.05a
    • /
    • pp.17-28
    • /
    • 2004
  • As Korean ginseng is hybrid, an individual variation is very severe, and it takes long times in new breeding because it is required 4 years to pick the seed. But, transformation technique makes the high-functional breeding in short time. The focus of these ginseng studies is to find and secure the useful gene. And it is urgent to accumulate the fundamental data for the molecular breeding and secure the useful genes. Therefore, transformation and soil acclimatization technique are necessary to molecular breeding in use of the introduction of functional genes. In this study, it add to secure of new regulation gene and useful gene as to accumulate the fundamental data for the place where it will contribute to raise the national competitive power. To analyze the useful genes in large scale, we constructed CDNA libraries with various tissues, species, and treated tissue. EST analysis of ginseng perform in large scale and build the EST database of ginseng. We perform the full length sequencing about the selected lots of clones that include the entire open reading frame of the amino acid residues and construct cDNA chip with the parental EST clones. Establishment of the transformation and a soil acclimatization system throuth the re-introduction of the selected ginseng gene that related with the secondary metabolism and anti-stress into the ginseng.

  • PDF

Calmodulin of Olive Flounder Paralichthys olivaceus : Cloning and Expression Analysis

  • Hong, Gyeong-Eun;Kong, Hee Jeong;Nam, Bo-Hye;Kim, Young-Ok;Kim, Woo-Jin;Lee, Sang-Jun;Choi, Tae-Jin
    • Journal of Marine Bioscience and Biotechnology
    • /
    • v.2 no.4
    • /
    • pp.234-237
    • /
    • 2007
  • Calmodulin (CaM) is a $Ca^{2+}$-binding protein essential for biological functions mediated through $Ca^{2+}$-dependent mechanism. A cDNA clone for CaM was isolated from a cDNA library of olive flounder Paralichthys olivaceus. The CaM cDNA concists of 782 bp and encodes a polypeptide of 149 amino acids with four $Ca^{2+}$-binding motifs EF-hands (EF-I, EF-II, EF-III, and EF-IV). The deduced amino acid sequence of CaM shows 97-100% amino acid sequence identity to other CaM sequences. Semi-quantitative PCR analysis revealed that the CaM transcription was began during early development and the CaM mRNA is expressed highly in brain and intestine, and moderately in kidney, gill, and eye of healthy olive flounder. Taken together, CaM may be necessary for early olive flounder development and that it may have a part in homeostasis.

  • PDF

Molecular Cloning, Sequence Analysis, and in Vitro Expression of Flavanone 3β-Hydroxylase from Gypsophila paniculata (안개초(Gyposphila paniculata)로부터 Flavanone 3β-Hydroxylase 유전자의 분리 및 분석)

  • Min, Byung-Whan
    • Journal of Plant Biotechnology
    • /
    • v.33 no.2
    • /
    • pp.85-91
    • /
    • 2006
  • Flavanone 3$\beta$-hydroxylase (FHT) is an enzyme acting in the central part of the flavonoid biosynthesis pathway. FHT catalyses the hydroxylation of flavanone to dihydroflavonols in the anthocyanin pathway. In this paper we describe the cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme FHT in Gypsophila paniculata L. A heterologous cDHA probe from Dianthus cavophyllus was used to isolate FHT-encoding cDHA clones from Gypsophila paniculata L.. Inspection of the 1471 bp long sequence revealed an open reading frame 1047 bp, including a 190 bp 5' leader region and 288 bp 3' untranslated region. Comparison of the coding region of this FHT cDHA sequence including the sequences of Arabidopsis thaliana, Citrus sinensis, Dianthus caryophyllus, Ipomoea batatas, Matthiola incana, Nierembergia sp, Petunia hybrida, Solanum tuberosum, Vitis vinifera reveals a identity higher than 69% at the nucleotide level. The function of this nucleotide sequences were verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRHA from wild type and mutant plants, by in vitro expression yielding and enzymatically active hydroxylase, as indicated by the small dihydrokaempferol peak. Genomic southern blot analysis showed the presence of only one gene for FHT in Gypsophila paniculata.

Distance measurement System from detected objects within Kinect depth sensor's field of view and its applications (키넥트 깊이 측정 센서의 가시 범위 내 감지된 사물의 거리 측정 시스템과 그 응용분야)

  • Niyonsaba, Eric;Jang, Jong-Wook
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
    • /
    • 2017.05a
    • /
    • pp.279-282
    • /
    • 2017
  • Kinect depth sensor, a depth camera developed by Microsoft as a natural user interface for game appeared as a very useful tool in computer vision field. In this paper, due to kinect's depth sensor and its high frame rate, we developed a distance measurement system using Kinect camera to test it for unmanned vehicles which need vision systems to perceive the surrounding environment like human do in order to detect objects in their path. Therefore, kinect depth sensor is used to detect objects in its field of view and enhance the distance measurement system from objects to the vision sensor. Detected object is identified in accuracy way to determine if it is a real object or a pixel nose to reduce the processing time by ignoring pixels which are not a part of a real object. Using depth segmentation techniques along with Open CV library for image processing, we can identify present objects within Kinect camera's field of view and measure the distance from them to the sensor. Tests show promising results that this system can be used as well for autonomous vehicles equipped with low-cost range sensor, Kinect camera, for further processing depending on the application type when they reach a certain distance far from detected objects.

  • PDF

Sequencing of cDNA Clones Expressed in Adipose Tissues of Korean Cattle

  • Bong, J.J.;Tong, K.;Cho, K.K.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.18 no.4
    • /
    • pp.483-489
    • /
    • 2005
  • To understand the molecular mechanisms that regulate intramuscular fat deposition and its release, cDNA clones expressed in adipose tissues of Korean cattle were identified by differential screening from adipose tissue cDNA library. By partial nucleotide sequencing of 486 clones and a search for sequence similarity in NCBI nucleotide databases, 245 clones revealed unique clones. By a functional grouping of the clones, 14% of the clones were categorized to metabolism and enzyme-related group (stearoyl CoA desaturase, lactate dehydrogenase, fatty acid synthase, ATP citrate lyase, lipoprotein lipase, acetyl CoA synthetase, etc), and 6% to signal transduction/cell cycle-related group (C/EBP, cAMP-regulated phosphoprotein, calmodulin, cyclin G1, cyclin H, etc), and 4% to cytoskeleton and extracellular matrix components (vimentin, ankyrin 2, gelosin, syntenin, talin, prefoldin 5). The obtained 245 clones will be useful to study lipid metabolism and signal transduction pathway in adipose tissues and to study obesity in human. Some clones were subjected to full-sequencing containing open reading frame. The cDNA clone of bovine homolog of human prefoldin 5 gene had a total length of 959 nucleotides coding for 139 amino acids. Comparison of the deduced amino acid sequences of bovine prefoldin 5 with those of human and mouse showed over 95% identity. The cDNA clone of bovine homolog of human ubiquitin-like/S30 ribosomal fusion protein gene had a total length of 484 nucleotides coding for 133 amino acids. Comparison of the deduced amino acid sequences of bovine ubiquitin-like/S30 ribosomal fusion protein gene with those of human, rat and mouse showed over 97% identity. The cDNA clone of bovine homolog of human proteolipid protein 2 mRNA had a total length of 928 nucleotides coding for 152 amino acids. Comparison of the deduced amino acid sequences of bovine proteolipid protein 2 with those of human and mouse showed 87.5% similarity. The cDNA clone of bovine homolog of rat thymosin beta 4 had a total length of 602 nucleotides coding for 44 amino acids. Comparison of the deduced amino acid sequences of bovine thymosin beta 4 gene with those of human, mouse and rat showed 93.1% similarity. The cDNA clone of bovine homolog of human myotrophin mRNA had a total length of 790 nucleotides coding for 118 amino acids. Comparison of the deduced amino acid sequences of bovine myotrophin gene with those of human, mouse and rat showed 83.9% similarity. The functional role of these clones in adipose tissues needs to be established.

Molecular Cloning of the cDNA of Heat Shock Protein 88 Gene from the Entomopathogenic Fungus, Paecilomyces tenuipes Jocheon-1

  • Liu, Ya-Qi;Park, Nam Sook;Kim, Yong Gyun;Kim, Keun Ki;Park, Hyun Chul;Son, Hong Joo;Hong, Chang Ho;Lee, Sang Mong
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • v.28 no.2
    • /
    • pp.71-84
    • /
    • 2014
  • The full-length heat shock protein 88 (HSP88) complementary DNA (cDNA) of Paecilomyces tenuipes Jocheon-1 was obtained by screening the Paecilomyces tenuipes (P. tenuipes) Jocheon-1 Uni-Zap cDNA library and performing 5' RACE polymerase chain reaction (PCR). The P. tenuipes Jocheon-1 HSP88 cDNA contained an open reading frame (ORF) of 2,139-basepair encoding 713 amino acid residues. The deduced amino acid sequence of the P. tenuipe s Jocheon-1 HSP88 cDNA showed 77% identity to Nectria haematococca HSP88 and 45-76% identity to other fungal homologous HSP88s. Phylogenetic analysis and BLAST program analysis confirmed that the deduced amino acid sequences of the P. tenuipes Jocheon-1 HSP88 gene belonged to the ascomycetes group within the fungal clade. The P. tenuipes Jocheon-1 HSP88 also contained the conserved ATPase domain at the N-terminal region. The cDNA encoding P. tenuipes Jocheon-1 HSP88 was expressed as an 88 kilodalton (kDa) polypeptide in baculovirus-infected insect Sf9 cells. Under higher temperature conditions for the growth of the entomopathogenic fungus, mRNA expression of P. tenuipes Jocheon-1 HSP88 was quantified by real time PCR (qPCR). The results showed that heat shock stress induced a higher level of mRNA expression compared to normal growth conditions.