Application of two-dimensional protein analysis for strain fingerprinting and mutant analysis of Azospirillum species

1989 ◽  
Vol 35 (10) ◽  
pp. 960-967 ◽  
Author(s):  
René De Mot ◽  
Jos Vanderleyden
Keyword(s):  
Gene Dosage ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein Analysis ◽  
Resolving Power ◽  
Two Dimensional ◽  
Wild Type ◽  
Protein Patterns ◽  
Gene Dosage Effect ◽  
Chemically Induced ◽  
Mutant Strains

Phenol-extracted total proteins from wild-type and mutant strains of Azospirillum brasilense and A. lipoferum were subjected to two-dimensional polyacrylamide gel electrophoresis. The protein patterns of both species could be readily distinguished by specific configurations of a limited number of spots. The resolving power of the technique allowed its application for strain fingerprinting. Thus, near-identity of A. brasilense strains Sp7 and Cd was demonstrated. In addition, minor changes in protein profiles resulting from spontaneous, chemically induced, or transposon-mediated mutations in the A. brasilense genome were evidenced. For a nitrate reductase negative mutant of strain Sp245 and a Sp7 mutant affected in exopolysaccharide synthesis, production of a truncated protein was involved. In Tn5-generated mutants, a gene dosage effect for the transposon-encoded neomycine phosphotransferase was observed.Key words: Azospirillum, two-dimensional protein analysis, fingerprinting.

scholarly journals Two-dimensional Gel Analysis of Proteins from Mouse Fetuses with Trisomy 19 after DEAE-Sepharose Chromatography

1986 ◽  
Vol 47 (3) ◽  
pp. 193-197 ◽  
Author(s):  
Gerd H. Reichert
Keyword(s):  
Gene Dosage ◽  
Protein Pattern ◽  
Polyacrylamide Gel Electrophoresis ◽  
Two Dimensional ◽  
Protein Patterns ◽  
Gene Dosage Effect ◽  
Gene Dosage Effects ◽  
The Individual ◽  
Mouse Fetuses ◽  
Primary Gene

SummaryIsoelectrofocusing two-dimensional polyacrylamide gel electrophoresis (IEF-2D-PAGE) offers the opportunity to detect typical alterations in the protein pattern of trisomic mouse foetuses at a given time of development. The fractionation of the cell lysate by differential centrifugation into various subcellular components (nuclei, membranes, polyribosomes, cytoplasmic proteins) and fractionation of the proteins through DEAE-Sepharose chromatography allows detection of protein differences.It is possible to detect eight differences in the protein patterns between trisomy 19 (Ts 19) mouse foetuses and euploid mouse fetuses at day 15. Five of these differences are quantitative in nature, three are qualitative. One of these proteins is synthesized in Ts 19 foetuses at a higher level than in euploid mouse fetuses (primary gene dosage effect). The other seven proteins are reduced or not present in trisomic foetuses (consequences of primary gene dosage effects).The molecular mass of the individual proteins ranges from 13 to 41 kDa.

Variation in goat fibre protein

1989 ◽  
Vol 40 (3) ◽  
pp. 675 ◽  
Author(s):  
DJ Tucker ◽  
AHF Hudson ◽  
A Laudani ◽  
RC Marshall ◽  
DE Rivett
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Apparent Molecular Weight ◽  
Wool Fibre ◽  
Two Dimensional ◽  
Protein Patterns ◽  
Cashmere Goats

The proteins from a range of cashmere, mohair, angoratcashmere crossbred and wool fibre samples were extracted at pH 8 with 8 M urea containing dithiothreitol, and were then radiolabelled by S-carboxymethylation using iodo(2-14C) acetate. The proteins from each sample were examined by two dimensional polyacrylamide gel electrophoresis in which the separation in the first dimension was according to charge at pH 8.9 and in the second dimension according to apparent molecular weight in the presence of sodium dodecyl sulfate. After electrophoresis the proteins were detected by fluorography. Protein differences in keratin samples from some individual goats existed, although the overall protein patterns were similar. None of the differences were consistent with any one goat fibre type. The protein patterns obtained for fibre samples from individual cashmere goats showed some differences when compared to those found for commercial blends from the same country of origin, indicating that blending can mask any animal-to-animal variation. While the electrophoretic technique does not unequivocally distinguish between cashmere, mohair and angora/cashmere crossbred fibres it does differentiate between wool and goat fibres.

scholarly journals Impact of gene dosage, loss of wild-type allele, and FLT3 ligand on Flt3-ITD–induced myeloproliferation

Blood ◽  
2011 ◽  
Vol 118 (13) ◽  
pp. 3613-3621 ◽  
Author(s):  
Shabnam Kharazi ◽  
Adam J. Mead ◽  
Anna Mansour ◽  
Anne Hultquist ◽  
Charlotta Böiers ◽  
...  
Keyword(s):  
Cancer Progression ◽  
Gene Dosage ◽  
Growth Factor Receptor ◽  
Wild Type Allele ◽  
Flt3 Ligand ◽  
Wild Type ◽  
Gene Dosage Effect ◽  
Internal Tandem Duplication ◽  
Type Allele ◽  
Tandem Duplications

Abstract Acquisition of homozygous activating growth factor receptor mutations might accelerate cancer progression through a simple gene-dosage effect. Internal tandem duplications (ITDs) of FLT3 occur in approximately 25% cases of acute myeloid leukemia and induce ligand-independent constitutive signaling. Homozygous FLT3-ITDs confer an adverse prognosis and are frequently detected at relapse. Using a mouse knockin model of Flt3–internal tandem duplication (Flt3-ITD)–induced myeloproliferation, we herein demonstrate that the enhanced myeloid phenotype and expansion of granulocyte-monocyte and primitive Lin−Sca1+c-Kit+ progenitors in Flt3-ITD homozygous mice can in part be mediated through the loss of the second wild-type allele. Further, whereas autocrine FLT3 ligand production has been implicated in FLT3-ITD myeloid malignancies and resistance to FLT3 inhibitors, we demonstrate here that the mouse Flt3ITD/ITD myeloid phenotype is FLT3 ligand-independent.

MOUSE MUTANT DEFICIENT IN d-AMINO ACID OXIDASE ACTIVITY

Genetics ◽  
1983 ◽  
Vol 103 (2) ◽  
pp. 277-285 ◽  
Author(s):  
Ryuichi Konno ◽  
Yosihiro Yasumura
Keyword(s):  
Enzyme Activity ◽  
Amino Acid ◽  
Oxidase Activity ◽  
Null Allele ◽  
Gene Dosage ◽  
Autosomal Gene ◽  
Acid Oxidase ◽  
Wild Type ◽  
Amino Acid Oxidase ◽  
Gene Dosage Effect

ABSTRACT d-Amino acid oxidase activity in the kidney homogenates of mice of seven strains was measured to search for a mutant for this enzyme. There was a consistent sex difference in the enzyme activity in these strains: male mice showed higher levels of the enzyme activity than females. In contrast to other strains, some mice of the ddY strain did not possess enzyme activity. This trait was inheritable, and a mouse stock without enzyme activity (DAO-) was established. The allele (Dao-1c) carried by the DAO- mice was recessive and behaved as a single autosomal gene in inheritance. Heterozygous mice for this gene (Dao-1  +/Dao-1c) showed nearly half the enzyme activity of the wild-type homozygotes (Dao-1  +/Dao-1  +), suggesting that Dao-1c is a null allele and that there is a gene dosage effect on the enzyme activity.

scholarly journals VanA-Type Staphylococcus aureus Strain VRSA-7 Is Partially Dependent on Vancomycin for Growth

2009 ◽  
Vol 53 (9) ◽  
pp. 3657-3663 ◽  
Author(s):  
Carole Moubareck ◽  
Djalal Meziane-Cherif ◽  
Patrice Courvalin ◽  
Bruno Périchon
Keyword(s):  
Staphylococcus Aureus ◽  
Gene Dosage ◽  
Conjugative Plasmid ◽  
Aureus Strain ◽  
Wild Type ◽  
Two Component System ◽  
Gene Dosage Effect ◽  
Peptidoglycan Synthesis ◽  
Resistance Pathway ◽  
Two Component

ABSTRACT VanA-type Staphylococcus aureus strain VRSA-7 was partially dependent on glycopeptides for growth. The vanA gene cluster, together with the erm(A) and the ant(9)-Ia resistance genes, was carried by the ca. 35- to 40-kb conjugative plasmid pIP848 present at five copies per cell. The chromosomal ddl gene had a mutation that led to a N308K substitution in the d-Ala:d-Ala ligase that resulted in a 1,000-fold decrease in activity relative to that of strain VRSA-6. Strain VRSA-7 grown in the absence or in the presence of vancomycin mainly synthesized precursors ending in d-Ala-d-Lac, indicating that the strain relied on the vancomycin resistance pathway for peptidoglycan synthesis. Greatly enhanced growth in the presence of glycopeptides and the absence of mutations in the genes for VanR and VanS indicated the inducible expression of resistance. Thus, a combination of loose regulation of the vanA operon by the two-component system and a gene dosage effect accounts for the partial glycopeptide dependence of VRSA-7. Since peptidoglycan precursors ending in d-Ala-d-Lac are not processed by PBP 2′, the strain was fully susceptible to oxacillin, despite the production of a wild-type PBP 2′.

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