• Title/Summary/Keyword: gene library

Search Result 666, Processing Time 0.028 seconds

Molecular Cloning and Analysis of Sporulation-Specific Glucoamylase (SGA) Gene of Saccharomyces diastaticus

  • Kang, Dae-Ook;Hwang, In-Kyu;Oh, Won-Keun;Lee, Hyun-Sun;Ahn, Soon-Cheol;Kim, Bo-Yeon;Mheen, Tae-Ick;Ahn, Jong-Seog
    • Journal of Microbiology
    • /
    • v.37 no.1
    • /
    • pp.35-40
    • /
    • 1999
  • Sporulation-specific glucoamylase (SGA) gene was isolated from genomic library of Saccharomyces diastaticus 5114-9A by using glucoamylase non-producing mutant of S. diastaticus as a recipient. When the glucoamylase activities of culture supernatant, periplasmic, and intracellular fraction of cells transformed with hybrid plasmid containing SGA gene were measured, the highest activity was detected in culture supernatant. SGA produced by transformant and extracellular glucoamylase produced by S. diastaticus 5114-9A differed in enzyme characteristics such as optimum temperature, thermostability, and resistance to SDS and urea. But the characteristics of SGA produced by sporulating yeast cells and vegetatively growing transformants were identical.

  • PDF

Bacterial Diversity at Different Depths in Lead-Zinc Mine Tailings as Revealed by 16S rRNA Gene Libraries

  • Zhang, Han-Bo;Shi, Wen;Yang, Ming-Xia;Sha, Tao;Zhao, Zhi-Wei
    • Journal of Microbiology
    • /
    • v.45 no.6
    • /
    • pp.479-484
    • /
    • 2007
  • Bacterial communities at 10 cm, 100 cm, and 200 cm depths in a 100-year-old lead-zinc tailing heap were evaluated by constructing 16S rRNA gene libraries. In total, 98 operational taxonomic units (OTUs) were identified from 193 clones at a 3% sequence difference level. The OTU number and species richness decreased with the depth. Species composition was significantly different between the three libraries. Fifty-seven percent of the examined clones were Acidobacteria and 27% belonged to Proteobacteria. Other sequences included Chloroflexi, Firmicutes, Chlamydiae, Actinobacteria, Gemmatimonadetes, Nitrospira, and three unclassified OTUs. Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria were mainly distributed in the rhizosphere of naturally colonizing plants; however, Deltaproteobacteria, Acidobacteria, and Chloroflexi tended to inhabit the deeper tailings (below the 100 cm-depth).

Nucleotide Sequence Analysis and Expression of the Alginate Lyase Gene from Pseudomonas sp. W7 in Escherichia coli

  • Lee, Jong-Hee;Kang, Jung-Hwa;Kim, Young-Ok;Kim, Jin-Man;Kong, In-Soo
    • Journal of Microbiology and Biotechnology
    • /
    • v.8 no.5
    • /
    • pp.531-535
    • /
    • 1998
  • The gene encoding alginate lyase was isolated from a library constructed with the vector, pUC19, and expressed in Escherichia coli. The nucleotide sequence of the cloned alginate lyase gene (ALY) from Pseudomonas sp. W7 was determined. The nucleotide sequence revealed a 1,035 bp open reading frame (ORF), encoding 345 amino acid residues with a calculated molecular mass of 37,478 Da. The N-terminal amino acid sequences (15 residues) of purified alginate lyase corresponded to that of the deduced amino acid sequence.

  • PDF

Isolation of Novel Pseudonocardia Polyene Biosynthetic Genes via Genomics-based PCR Screening

  • Lee, Mi-Yeon;Hwang, Young-Bin;Park, Hyun-Joo;Han, Kyu-Boem;Kim, Eung-Soo
    • 한국생물공학회:학술대회논문집
    • /
    • 2005.04a
    • /
    • pp.396-397
    • /
    • 2005
  • The polyene antibiotics are a family of most promising antifungal polyketide compounds, typically produced by actinomycetes species. Using the polyene CYP-specific PCR screening with served actinomycetes genomic DNAs, Pseudonocardia autotrophica strain was identified to contain a unique polyene-specific CYP gene. The genomic DNA library screening using the polyene-specific CYP gene probe revealed the positive cosmid clone containing an approximately 34.5 kb DNA fragment revealed a total of seven complete and two incomplete open reading frame (ORFs), which are highly homologous but unique to previously-known polyene biosynthetic genes. These results suggest that the polyene-specific screening approach should be an efficient way of isolating potectially-valuable cryptic polyene biosynthetic gene cluster from various rare actinomycetes.

  • PDF

Effects of 17 β -estradiol, bisphenol A and genistein on the expression of the glutathione peroxidase gene of Philasterides dicentrarchii (Ciliophora: Scuticociliata)

  • Lee, Eun-Hye;Kim, Sung-Mi;Nam, Yoon-Kwon;Kim, Ki-Hong
    • Journal of fish pathology
    • /
    • v.19 no.3
    • /
    • pp.189-195
    • /
    • 2006
  • A subtracted cDNA library of a marine scuticociliate, Philasterides dicentrarchii, in response to 17β-estradiol exposure was constructed using suppression subtractive hybridization (SSH). As a result of SSH, 275 clones were isolated, and among them, only glutathione peroxidase (GPX) gene was isolated as an antioxidative enzyme responding to 17β-estradiol. The semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis revealed that the transcription of GPX gene of P. dicentrarchii was clearly increased by exposure to 17β-estradiol. The GPX transcription was also clearly increased by exposure to xenoestrogens such as bisphenol A (BPA) and genistein.

SEQUENCE ANALYSIS AND COMPARISON OF BOVINE αS1-CASEIN GENOMIC DNA

  • Lin, C.S.;Huang, M.C.;Choo, K.B.;Tseng, Y.H.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.6 no.4
    • /
    • pp.541-547
    • /
    • 1993
  • A phage clone containing the partial ${\alpha}_{S1}$-casein gene was isolated from a bovine genomic library by using mixed probes of ovine ${\alpha}_{S1}$-, ${\beta}$- and ${\kappa}$-casein cDNAs. Restriction enzyme mapping analysis for 14.6 kb revealed that the map was in conflict with the report of Meade et al. (1990), especially in the 3'-end fragment. Sequence analysis of 12.6 kb revealed a high AT/GC ratio (1.64); we have identified eight exon sequences according to the bovine ${\alpha}_{S1}$-casein cDNA sequence. The same exon/intron splice junction sequence was observed between these exons. We suggest that the bovine ${\alpha}_{S1}$-casein gene night contain a minimum of 18 exons and the full length is approximately 18-19 kb.

Cloning of CAB cDNA encoding chlorophyll a/b binding protein of photosystem II in Korean ginseng and Use in Plant (고려인삼 광계 II Chlorophyll a/b binding Protein 유전자(CAB)의 cloning 및 식물에의 활용연구)

  • 김갑식;이기원;이종철;여운형;채순용;박은경
    • Journal of the Korean Society of Tobacco Science
    • /
    • v.21 no.2
    • /
    • pp.152-159
    • /
    • 1999
  • A CAB cDNA clone(pKGCAB) encoding the light harvesting chlorophyll a/b binding protein of the semi-shade plant, Korean ginseng(Panax ginseng C. A. Meyer) was isolated by the one-way path random sequencing of ginseng cDNA library clones and transgenic tobacco plants(Nicotiana tabacum NC82) were produced by the transformation of this ginseng CAB gene in use of Agrobacterium tumefaciens LBA4404. The CAB gene showed type 1 structure of LHCP-II, 84% similarity in nucleotide sequence and 92% in amino acid sequence to that of Nicotiana tabacum CAB40, respectively. Seed germination and initial growth of the transgenic tobacco plants transformed with the cDNA fragment were accelerated under low light intensity compared with those of normal tobacco plant, that may result from the higher light sensitivity of the transgenic plants than that of the normal.

  • PDF

Molecular Cloning of the cCDNA for $NAD^+$ /-dependent 15-hydroxyprostagladin Dehydrogenase Gene Homologue from the Mole Cricket, Gryllotalpa orientalis

  • Kim, Iksoo;Lee, Young-Sin;Kim, Eun-Sun;Lee, Heui-Sam;Kim, Jin-Won;Ahn, Mi-Young;Ryu, Kang-Sun
    • Proceedings of the Korean Society of Sericultural Science Conference
    • /
    • 2003.04a
    • /
    • pp.67-67
    • /
    • 2003
  • The NAD$^{+}$-dependent 15-hydroxyprostagladin dehydrogenase (15-PGDH) is a key catabolic enzyme responsible for the control of the biological activities of prostagladins. So far the gene has been found in a diverse organism including three insect dipteran species and one lepidopteran species. In this study, a cDNA encoding the 15-PGDH gene homologue was isolated from the cDNA library of the mole cricket, Gryllotalpa orientalis. (omitted)d)

  • PDF

Initial Transcriptome Profile of Rainbow Trout (Oncorhynchus mykiss) Liver

  • Kim Soonhag
    • Fisheries and Aquatic Sciences
    • /
    • v.6 no.1
    • /
    • pp.41-44
    • /
    • 2003
  • Ninety nine random complementary DNA clones from rainbow trout (Oncorhynchus mykiss) liver cDNA library were partially sequenced as one approach to analyze the transcribed sequences of its genome. Of the sequence generated, $64.0\%$ of the ESTs were represented by 29 known genes. Thirty six clones of the unknown gene products potentially represent 31 unique genes. Serum albumin $(16.1\%)$ was the most abundant in the liver. The structural genes in the liver $(19\%)$ were the highly expressed functional category. This research is helpful to understand tissue specific gene expression profile and basic relationship between tissue and functional categories of the genes.

Nucleotide Sequence of a Proteinase Inhibitor I Gene in Potato (감자에 존재하는 단백질분해효소 억제제 I 유전자의 염기서열)

  • 이종섭
    • Journal of Plant Biology
    • /
    • v.32 no.2
    • /
    • pp.67-78
    • /
    • 1989
  • Hybridization of DNA isolated from leaves of Russet Burbank potato with tomato cDNA as a probe revealed the presence of about ten inhibitor 1 genes in the genome. Screening of a genomic library of Russet Burbank potato resulted in isolation of seven different genomic clones carrying inhibitor I genes. One of the genomic clones, clone 2, contained two EcoRI fragments of 3.4 and 1.8 kb in size, respectively, which were hybridized with the probe. The nucleotide sequence of parts of the hybridizing EcoRI fragments revealed that they contain a complete gene which codes for an open reading frame of 107 amino acids. It is interrupted by two intervening sequences of 502 and 493 bp, situated at the positions of codons 17 and 43, respectively, of the open reading frame. Putative regulatory sequences, TATAAA and CCACT, were found at the 5' flanking region. In addition, a copy of a 100 bp repeat found at a tomato inhibitor I gene was identified.

  • PDF