• Title/Summary/Keyword: Gene characterization

Search Result 2,030, Processing Time 0.034 seconds

The Structural Characterization of the Putative DNA-Binding Protein BldB from Streptomyces Lividans

  • Ochiriin, Tsogbadrakh-Mishig;Kang, Sa-Ouk
    • Proceedings of the Korean Biophysical Society Conference
    • /
    • 2002.06b
    • /
    • pp.49-49
    • /
    • 2002
  • Mutants blocked at the earliest stages of morphological development in Streptomyces species are called bld mutants. We have cloned bldB gene ORF from Slividans. Genomic Southern blot analysis for main strains S.lividans, S.seoulensis, S.coelicolor A3(2), and S.griseus indicated that bldB gene is conserved in all main Streptomyces strains.(omitted)

  • PDF

Characterization of PpsR, a Transcriptional Repressor of the Expression of Photosysem Gene, from Rhodobacter sphaeroides

  • Cho, Seung-Hyun;Kang, Sa-Ouk
    • Proceedings of the Korean Biophysical Society Conference
    • /
    • 2002.06b
    • /
    • pp.51-51
    • /
    • 2002
  • PpsR from the facultative photohetrotroph Rhodobacter sphaeroides is involved in repression of photo system gene expression. SDS-PAGE analysis showed that some portion of PpsR is oxidized so that intra- or inter-disulfide bond is formed between the two cysteins in each subunit. The disulfide bond was reduced by dithiothreitol and the binding activity to puc promoter region was increased.(omitted)

  • PDF

Characterization of the recombinant cellulase A from Thermotoga maritima

  • Kim, Chung Ho
    • Journal of Applied Biological Chemistry
    • /
    • v.64 no.3
    • /
    • pp.213-216
    • /
    • 2021
  • A gene encoding thermostable cellulase A (TmCelA) was isolated from Thermotoga maritima. The open reading frame of TmCelA gene was 774 bp long which predicted to encode 257 amino acid residues with a molecular weight of 29,732 Da. To examine the biochemical properties, the TmCelA was overexpressed in E. coli BL21, and expressed protein was purified. The optimum temperature of recombinant TmCelA was 90-95 ℃, and the optimum pH of recombinant TmCelA was approximately pH 5.0. Recombinant TmCelA was stable at temperature below 90 ℃.

Expression and Characterization of Escherichia coli Adhesin Protein Linked to Cholera Toxin A2/B Subunits in Escherichia coli

  • Lee, Young-Hwa;Ryu, Dong-Kyun;Rhee, Dong-Kwon;Pyo, Suhk-Neung
    • Proceedings of the PSK Conference
    • /
    • 2002.10a
    • /
    • pp.309.2-309.2
    • /
    • 2002
  • The FimH subunit of type l-fimbriated Escherichia coli has been determined as a major cause of urinary tract infection. To produce a possible vaccine antigen against urinary tract infection, the fimH gene was genetically coupled to the ctxa2b gene, which was then cloned into pMAL -p2E expression vector. The chimaeric construction of pMALfimH/ctxa2b was transformed into Escherichia coli TB1 and its N-terminal amino acid sequence was analyzed. (omitted)

  • PDF

Cloning. Sequencing and Characterization of the Urease Gene Cluster of the Streptococcus vestibularis

  • Kim, Geun-Y.;Lee, Mann-H.
    • Proceedings of the PSK Conference
    • /
    • 2002.10a
    • /
    • pp.332.1-332.1
    • /
    • 2002
  • Streptococcus vestibularis is a urease-producing oral bacterium. frequently isolated from vestibular mucosa of human oral cavity. Ureolysis by S. vestibularis and other ureolytic oral bacteria is believed to be crucially involved in oral microbial ecology and oral health. Genomic library of the S. vestibularis ATCC49124 was constructed in an E. coli plasmid vector and the urease-positive transformants harboring the urease gene cluster were isolated on Christensen-urea agar plates. The minimal DNA region required for the urease activity was located on a 5.6 kb DNA fragment. (omitted)

  • PDF

Positional Cloning and Phenotypic Characterization of a New Mutant Mouse with Neuronal Migration Abnormality

  • Park, Chankyu;Ackerman, Susan-L
    • Proceedings of the Korean Society of Developmental Biology Conference
    • /
    • 2001.10a
    • /
    • pp.14-17
    • /
    • 2001
  • Positional clonging (map-based cloning) of mutations or genetic variations has been served as an invaluable tool to understand in-vivo functions of genes and to identify molecular components underlying phenotypes of interest. Mice homozygous for the cerebellar deficient folia (cdf) mutation are ataxic, with cerebellar hypoplasia and abnormal lobulation of the cerebellum. In the cdf mutant cerebellum approximately 40% of Purkinje cells are ectopically located within the white matter and the inner granule cell layer (IGL). To identify the cdf gene, a high-resolution genetic map for the cdf-gene-encompassing region was constructed using 1997 F2 mice generated from C3H/HeSnJ-cdf/cdf and CAST/Ei intercross. The cdf gene showed complete linkage disequilibrium with three tightly linked markers D6Mit208, D6Mit359, and D6Mit225. A contig using YAC, BAC, and P1 clones was constructed for the cdf critical region to identify the gene. A deletion in the cdf critical region on chromosome 6 that removes approximately 150 kb of DNA selection. cdf mutant mice with the transgenic copy of the identified gene restored the brain abnormalities of the mutant mice. The positional cloning of cdf gene provides a good example showing the identification of a gene could lead to finding a new component of important molecular pathways.

  • PDF

Cloning and Characterization of the $_L$-Lactate Dehydrogenase Gene (IdhL) from Lactobacillus reuteri ATCC 55739

  • Park, Jar-Yong;Park, Sun-Jung;Nam, Su-Jin;Ha, Yeong-Lae;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
    • /
    • v.12 no.5
    • /
    • pp.716-721
    • /
    • 2002
  • The ldhL gene encoding the $_L$-(+) lactate dehydrogenase was cloned from Lactobacillus reuteri ATCC 55739 chromosomal DNA and characterized. An internal 750-bp fiagment of ldhL gene was amplified by PCR using primers based on the conserved region of lactobacilli ldhL genes. A genomic library off. reuteri ATCC 55739 was constructed using pBR322, and colony hybridization experiments were performed using the 750-bp fragment as aprobe. One clone harboring a 4.0-kb PstI fragment was identified, and nucleotide sequencing confirmed it as an open reading frame of 972 bp in size in the middle. In addition to IdhL gene, an ORF homologous to Streptococcus pneumoniae TIGR4 hydrolase gene and 3' part of phosphomevalonate kinase gene (mvaK2) were also found on the 4 kb fragment. $_L$-LDH of L. reuteri ATCC 55739 showed the highest degree of homology with the $_L$-LDH of Pediococcus acidilactici (62.4%), fullowed by the $_L$-LDH of Lactobacillus pentosus (58.7%). The size of IdhL transcript determined by Northern blot was 1 kb, indicating the monocistronic nature of IdhL.

Genetic Characterization of the Escherichia coli O66 Antigen and Functional Identification of its wzy Gene

  • Cheng, Jiansong;Liu, Bin;Bastin David A.;Han, Weiqing;Wang, Lei;Feng Lu
    • Journal of Microbiology
    • /
    • v.45 no.1
    • /
    • pp.69-74
    • /
    • 2007
  • Escherichia coli is a clonal species, and occurs as both commensal and pathogenic strains, which are normally classified on the basis of their O, H, and K antigens. The O-antigen (O-specific polysaccharide), which consists of a series of oligosaccharide (O-unit) repeats, contributes major antigenic variability to the cell surface. The O-antigen gene cluster of E. coli O66 was sequenced in this study. The genes putatively responsible for the biosynthesis of dTDP-6-deoxy-L-talose and GDP-mannose, as well as those responsible for the transfer of sugars and for O-unit processing were identified based on their homology. The function of the wzy gene was confirmed by the results of a mutation test. Genes specific for E. coli O66 were identified via PCR screening against representatives of 186 E. coli and Shigella O type strains. The comparison of intergenic sequences located between galF and the O-antigen gene cluster in a range of E. coli and Shigella showed that this region may perform an important function in the homologous recombination of the O-antigen gene clusters.