Abstract:
:The use of plasmids possessing a regulatable gene coding for a site-specific recombinase together with its recognition sequences significantly facilitates genome manipulations since it allows self-excision of the portion of the genetic construct integrated into the host genome. Stable maintenance of such plasmids in Escherichia coli, which is used for plasmid preparation, requires prevention of recombinase synthesis in this host, which can be achieved by interrupting the recombinase gene with an intron. Based on this approach, Saccharomyces cerevisiae and Hansenula polymorpha self-excising vectors possessing intronated gene for Cre recombinase and its recognition sites (LoxP) were previously constructed. However, this work shows instability of the H. polymorpha vectors during plasmid maintenance in E. coli cells. This could be due to recombination between the loxP sites caused by residual expression of the cre gene. Prevention of translation reinitiation on an internal methionine codon completely solved this problem. A similar modification was made in a self-excising vector designed for S. cerevisiae. Apart from substantial improvement of yeast self-excising vectors, the obtained results also narrow down the essential part of Cre sequence.
journal_name
FEMS Microbiol Lettjournal_title
FEMS microbiology lettersauthors
Agaphonov MOdoi
10.1093/femsle/fnx222subject
Has Abstractpub_date
2017-12-01 00:00:00issue
22eissn
0378-1097issn
1574-6968pii
4563576journal_volume
364pub_type
杂志文章abstract::The transcription start point of the dnaK operon of Streptomyces coelicolor A3(2) has been determined by S1 mapping, using the EMBL automated fluorescent DNA sequencer. The -35 and -10 hexamers correspond to a sigma 70-type promoter. This promoter responds to heat shock and involves an inverted repeat different from t...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.1997.tb10327.x
更新日期:1997-04-15 00:00:00
abstract::The nucleotide sequence of the gene specifying the ethidium efflux system of Escherichia coli has been determined. The translated open reading frame has identified a membrane-bound polypeptide of 110 amino acids (11,960 Da) which shares 42% identity with a staphylococcal protein specifying resistance to ethidium. ...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1016/0378-1097(91)90338-b
更新日期:1991-08-01 00:00:00
abstract::To determine whether the luxA-luxB reporter system is suitable as a sensitive reporter for rapid real-time measurements of alpha-toxin gene (cpa) expression in Clostridium perfringens, and to widen the range of alpha-toxin-producing C. perfringens strains examined with respect to cpa expression during growth, the repo...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2000.tb09164.x
更新日期:2000-07-01 00:00:00
abstract::Two Pseudomonas aeruginosa genes encoding the enzymes 1-deoxy-D-xylulose 5-phosphate (DXP) synthase and DXP reductoisomerase, both involved in the mevalonate-independent biosynthesis of isoprenoids, have been expressed as recombinant enzymes in Escherichia coli. The purified P. aeruginosa DXP reductoisomerase was inhi...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2000.tb09307.x
更新日期:2000-09-15 00:00:00
abstract::Primers designed to conserved regions of botulinum and tetanus clostridial toxins were used to amplify DNA fragments from non-proteolytic Clostridium botulinum type F (202F) DNA using polymerase chain reaction technology. The fragments were cloned and the complete nucleotide sequence of the gene encoding type F toxin ...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1016/0378-1097(92)90408-g
更新日期:1992-09-15 00:00:00
abstract::We devised a screening method to obtain basidiomycetous fungi capable of degrading dioxins. About 200 fungal strains were selected from more than 1500 strains by their ability to decolorize Remazol brilliant blue R dye as an indicator. To attempt to eliminate the factor of dioxin sorption by mycelia, we prepared two s...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2002.tb11308.x
更新日期:2002-08-06 00:00:00
abstract::Peroxidase-conjugated transferrin was used to detect transferrin receptors both in intact outer membrane vesicles (OMVs) from Neisseria species in a dot blot assay, and in SDS-PAGE-separated OMV proteins after transferring to nitrocellulose membranes. All N. meningitidis strains produced transferrin receptors after cu...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.1991.tb04472.x
更新日期:1991-10-15 00:00:00
abstract::The effects of overexpression of the apurinic/apyrimidinic DNA endonuclease Nfo on wet and dry heat and UV-C (254 nm) resistance of Bacillus subtilis spores with or without alpha/beta-type small, acid-soluble spore proteins (SASP) were determined. Results revealed that overexpression of Nfo > or =50-fold in spores inc...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2009.01845.x
更新日期:2010-01-01 00:00:00
abstract::Staphylococcus aureus produces a number of virulence factors. The major virulence factors exhibited by S aureus include various antigens, enzymes, cytotoxins and exotoxins (e.g. hemolysins, enterotoxins and toxic shock syndrome toxin). In this report, we show the influence of punicalagin on the secretion of exoprotein...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1093/femsle/fnw253
更新日期:2016-11-01 00:00:00
abstract::Staphylococcus simulans biovar staphylolyticus secreted two bacteriolytic peptidoglycan hydrolases as proproteins that were activated as they were processed by an extracellular sulphydryl protease. This processing resulted in the production of multiple molecular-mass forms of each enzyme. Cells from early exponential ...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.1993.tb06321.x
更新日期:1993-06-15 00:00:00
abstract::The devR-devS two-component system of Mycobacterium tuberculosis was identified earlier and partially characterized in our laboratory. A devR::kan mutant of M. tuberculosis was constructed by allelic exchange. The devR mutant strain showed reduced cell-to-cell adherence in comparison to the parental strain in laborato...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1016/S0378-1097(04)00002-3
更新日期:2004-02-16 00:00:00
abstract::Listeria monocytogenes is a facultative intracellular pathogen responsible for both invasive and non-invasive food-borne illness in animals and humans. In this study, macrorestriction analysis following pulsed-field gel electrophoresis was used to show that Listeria monocytogenes serovar 1/2a strain EGD has a single c...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.1999.tb13632.x
更新日期:1999-06-15 00:00:00
abstract::The AbGst1 gene encoding a glutathione transferase from the necrotrophic pathogen Alternaria brassicicola was cloned from a benzyl isothiocyanate-treated conidial culture using differential display reverse transcription. The deduced amino-acid sequence of AbGst1p showed a significant degree of similarity to glutathion...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2006.00223.x
更新日期:2006-05-01 00:00:00
abstract::A lipopolysaccharide mutant of Leptospira interrogans (LaiMut) was obtained by growth in the presence of an agglutinating monoclonal antibody (mAb) against lipopolysaccharide. Agglutination reactions with anti-lipopolysaccharide mAbs and polyclonal antibodies showed that LaiMut had lost some serogroup Icterohaemorrhag...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2010.02029.x
更新日期:2010-08-01 00:00:00
abstract::Using natural rubber latex as the sole source of carbon and energy 50 rubber-degrading bacteria were isolated. Out of those 50 isolates, 33 were identified as Streptomyces species and 8 as Micromonospora species. Screening of 1220 bacteria obtained from different culture collections revealed 46 additional rubber-degra...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1016/s0378-1097(97)00072-4
更新日期:1997-05-15 00:00:00
abstract::The synthesis of short chain S-methyl thioesters was investigated in Geotrichum candidum strain GcG. The results indicated the involvement of an enzymatic reaction in this microorganism that led to the synthesis of S-methyl thioacetate (MTA) when methanethiol and acetyl-CoA were used as substrates. MTA was generated f...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2000.tb09430.x
更新日期:2000-12-15 00:00:00
abstract::We have studied the single rRNA gene cluster from the Archaeon, Pyrococcus furiosus. This isolate grows optimally at 100 degrees C and is thus a hyperthermophile. In P. furiosus, transcription of 16S rRNA is subject to regulation over a 7.5-fold range in response to a 20-fold increase in growth rate. The single cluste...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.1993.tb06379.x
更新日期:1993-08-01 00:00:00
abstract::We present a method to directly characterize the yeast diversity present in wine fermentations by employing denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR)-amplified 26S ribosomal RNA (rRNA) genes. PCR-DGGE of a portion of the 26S rRNA gene was shown to distinguish most yeast genera a...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2000.tb09210.x
更新日期:2000-08-01 00:00:00
abstract::SPC(3) is a multiple antigen peptide derived from the V(3) loop of human immunodeficiency virus (HIV) envelope (Env). It exerts a potent anti-HIV activity whereas it alters neither Env expression nor binding to CD(4). Here, SPC(3) binding characteristics, its subsequent intracellular fate and the fact that it inhibite...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2000.tb08964.x
更新日期:2000-02-15 00:00:00
abstract::Endpolygalacturonase I from Stereum purpureum has been identified as a causative substance for the silver-leaf disease in apples. It possesses a unique pro-sequence in the C-terminal region that lacks endpolygalacturonases from any other origin. In this study, we analyzed and compared enzymatic characteristics between...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1093/femsle/fnv134
更新日期:2015-09-01 00:00:00
abstract::We have constructed a very efficient synthesis and secretion system for cholera toxin B subunit (CTB) of Vibrio cholerae 569B using Bacillus-brevis. The constructed expression-secretion vector has the multiple promoters and the signal peptide coding region of the mwp gene, a structural gene for one of the major cell w...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.1993.tb06389.x
更新日期:1993-08-01 00:00:00
abstract::Among aerobic prokaryotes, many different terminal oxidase complexes have been described. Sequence comparison has revealed that the aa3-type cytochrome c oxidase and the bo3-type quinol oxidase are variations on the same theme: the heme-copper oxidase. A third member of this family has recently been recognized: the cb...
journal_title:FEMS microbiology letters
pub_type: 杂志文章,评审
doi:10.1111/j.1574-6968.1994.tb07067.x
更新日期:1994-08-01 00:00:00
abstract::A DNA-based method for the early detection and identification of decay basidiomycetes on wood chips is described. From 26 species of wood-decaying basidiomycetes and 20 species of wood-inhabiting ascomycetes, DNA fragments containing the internal transcribed spacers, 5.8S and partial 28S rDNA were amplified by PCR usi...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2002.tb11212.x
更新日期:2002-05-21 00:00:00
abstract::Bacterial swarming motility is a flagella-dependent translocation on the surface environment. It has received extensive attention as a population behavior involving numerous genes. Here, we report that Citrobacter freundii, an opportunistic pathogen, exhibits swarming movement on a solid medium surface with appropriat...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2011.02225.x
更新日期:2011-04-01 00:00:00
abstract::In this report we describe a Lactobacillus fermentum 3872 plasmid (pLF3872) not previously found in any other strain of this species. The analysis of the complete sequence of this plasmid revealed the presence of a gene encoding a large collagen-binding protein (CBP), as well as the genes responsible for plasmid maint...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1093/femsle/fnv068
更新日期:2015-06-01 00:00:00
abstract::A new Volvariella volvacea gene encoding a carbohydrate esterase (CE) family 4 acetyl xylan esterase (AXE) (designated as VvaxeII) was cloned and characterized. The coded polypeptide had 253 amino acid residues, with the first 19 serving as a secretion signal peptide. The VvaxeII transcript levels were high when the f...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2009.01585.x
更新日期:2009-06-01 00:00:00
abstract::Hopanoid fingerprints allowed to differentiate bacteria formerly connected to the genus Pseudomonas. Whereas all strains related to Pseudomonas and Ralstonia were devoid of any detectable hopanoid, these pentacyclic triterpenoids were found in the Burkholderia species and in related soil isolates, which contained as m...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2000.tb08974.x
更新日期:2000-02-15 00:00:00
abstract::Clostridium difficile, a Gram-positive spore-forming anaerobe, causes infections in humans ranging from mild diarrhoeal to potentially life-threatening pseudomembranous colitis. The availability of genomic information for a range of C. difficile strains affords researchers the opportunity to better understand not only...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.2010.02111.x
更新日期:2010-11-01 00:00:00
abstract::Chondroitin 6-sulfate depolymerizing activity was examined in the culture supernatant of Streptococcus intermedius ATCC 27335. 2-Acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-6-O-sulfo-D-galactose was split from the substrate. The enzyme(s) was not active upon chondroitin 4-sulfate or dermatan sulfate...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.1989.tb03284.x
更新日期:1989-01-15 00:00:00
abstract::DNA hybridization using the Synechocystis PCC6803 glucose transporter gene, glcP, revealed a single homologous region in two facultative photoautotrophic strains out of three tested, and none in three obligate autotrophs. In one of the latter, Synechococcus PCC7942, integration of glcP into the chromosome resulted in ...
journal_title:FEMS microbiology letters
pub_type: 杂志文章
doi:10.1111/j.1574-6968.1998.tb12959.x
更新日期:1998-04-15 00:00:00