Characterisation of high- and low-molecular-weight glutenin subunit genes in Chinese winter wheat cultivars and advanced lines using allele-specific markers and SDS-PAGE

2010 ◽  
Vol 61 (1) ◽  
pp. 84 ◽  
Author(s):  
F. P. Yang ◽  
L. H. Wang ◽  
J. W. Wang ◽  
X. Y. He ◽  
X. K. Zhang ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Molecular Markers ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Low Molecular Weight ◽  
Wheat Cultivars ◽  
Specific Pcr ◽  
Sds Page ◽  
Allele Specific ◽  
Chinese Wheat

Wheat end-use product quality is highly influenced by the composition and quantity of high- and low-molecular-weight glutenin subunits (HMW-GS and LMW-GS). In the present study, 224 Chinese wheat cultivars and advanced lines were characterised for the HMW-GS and LMW-GS with allele-specific PCR markers and sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that 56 cultivars (25.0%) carried the allele Glu-D1-1d (Dx5), while 80 cultivars (35.7%) with the allele Glu-B1-2a (By8) produced a 527-bp specific band. Fourteen genotypes (6.3%) with the allele Glu-B1e (Bx20) yielded a 701-bp amplicon with the marker Mar and a 753-bp specific PCR fragment with the marker ZSBy9aF1/R3. Glu-B1h (Bx14+By15) was present in only 1 genotype, and 2 cultivars contained the allele Glu-B1f (Bx13+By16) identified with the marker ZSBy9F2/R2. Four genotypes (1.8%) with the allele Glu-B1-1d (Bx6) gave 695-bp and 830-bp bands, and 5 genotypes (2.2%) with the allele Glu-B1i (Bx17+By18) amplified a 659-bp fragment using the marker Bx. One hundred and six cultivars (47.3%) had the allele Glu-B1-2b (By9), amplifying a 663-bp fragment with the marker ZSBy9aF1/R3; 34 genotypes (15.8%) contained the allele Glu-B3d, generating a 662-bp PCR fragment with the marker gluB3d. Fifteen cultivars (7.0%) with the allele Glu-B3b yielded 1570-bp and 750-bp PCR amplicons with the markers gluB3b and gluB3bef, respectively. The allele Glu-B3h was found in 7 cultivars, generating a 1022-bp PCR fragment with the marker gluB3h. The genotypes detected by SDS-PAGE were mostly consistent with those identified by molecular markers, confirming the utility of the molecular markers. The information for the HMW-GS and LMW-GS in Chinese wheat cultivars will be useful in wheat breeding programs.

Molecular detection of high- and low-molecular-weight glutenin subunit genes in common wheat cultivars from 20 countries using allele-specific markers

2011 ◽  
Vol 62 (9) ◽  
pp. 746 ◽  
Author(s):  
H. Jin ◽  
J. Yan ◽  
R. J. Peña ◽  
X. C. Xia ◽  
A. Morgounov ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecylsulfate ◽  
Functional Markers ◽  
Low Molecular Weight ◽  
Wheat Cultivars ◽  
High Frequencies ◽  
Allele Specific ◽  
End Use

The composition and quantity of high- and low-molecular-weight glutenin subunits (HMW-GS and LMW-GS) plays an important role in determining the end-use quality of wheat products. In the present study, 718 wheat cultivars and advanced lines from 20 countries were characterised for the HMW-GS and LMW-GS with allele-specific molecular markers. For the Glu-A1 locus, 311 cultivars (43.3%) had the subunit Ax2*, which predominated in cultivars from Canada (83.3%), Romania (91.7%), Russia (72.2%) and USA (72.2%). At Glu-B1 locus, 197 cultivars (27.4%) contained the By8 subunit and its frequency was higher in Japanese (60.0%) and Romanian (62.5%) genotypes than in those from other countries; 264 cultivars (36.8%) carried the By9 subunit, mostly existing in the cultivars from Austria (100.0%), Russia (72.2%), and Serbia (72.7%); the By16 subunit was present in 44 cultivars (6.1%), with a relatively high percentage in Chile (19.5%), whereas almost no cultivars from other countries had this subunit; the frequency of Bx7OE was 3.1%, and was found only in cultivars from Argentina (12.1%), Australia (4.1%), Canada (25.0%), Iran (20.0%), and Japan (30.0%). There were 446 genotypes (62.1%) with the subunit Dx5 at the Glu-D1 locus; high frequencies of Dx5 occurred in cultivars from Hungary (90.0%), Romania (95.8%), and Ukraine (92.3%). At the Glu-A3 locus, the frequencies of Glu-A3a, b, c, d, e, f and g were 2.9, 6.8, 53.2, 12.8, 7.7, 13.8, and 2.4%, respectively. Glu-A3a was detected only in the cultivars from Bulgaria (13.3%), China (12.2%), Germany (2.7%), Iran (6.7%), Mexico (14.3%), Turkey (4.7%), and USA (5.1%); the high frequencies of superior alleles Glu-A3b and d were found in cultivars from Australia (39.7%) and France (24.5%); Glu-A3c was widely distributed in cultivars from all the countries; the high frequencies of Glu-A3e, f and g were detected in cultivars from Argentina (33.3%), Canada (29.2%), and Hungary (20.0%). At the Glu-B3 locus, Glu-B3a, b, c, d, e, f, g, h and i were present in frequencies of 0.4, 22.3, 0.3, 2.8, 1.9, 3.9, 27.2, 18.8, and 7.1%, respectively. Glu-B3a was detected only in cultivars from Argentina (3.0%) and Ukraine (15.4%) cultivars; high frequencies of Glu-B3b and d were found in the cultivars from Romania (62.5%) and Mexico (14.3%); Glu-B3c was detected only in Romanian (8.3%) genotypes; frequencies of e, f, h and i were high in cultivars from Austria (40.0%), China (14.3%), USA (43.0%), and Argentina (33.3%); Glu-B3g was mostly detected in the cultivars from Germany (69.3%), Norway (77.3%), and Serbia (63.6%). The frequency of the 1B·1R translocation was 13.4%; it occurred in cultivars from all the countries except Australia, Austria, Norway, and Serbia. The functional markers applied in this study, in agreement with the results of sodium-dodecylsulfate–polyacrylamide gel electrophoresis, were accurate and stable, and can be used effectively in wheat quality breeding.

IDENTIFICATION OF CANADIAN AND AMERICAN WHEAT CULTIVARS BY SDS GRADIENT PAGE ANALYSIS OF GLIADIN AND GLUTENIN SUBUNITS

1987 ◽  
Vol 67 (4) ◽  
pp. 945-952 ◽  
Author(s):  
B. A. MARCHYLO
Keyword(s):  
Molecular Weight ◽  
Spring Wheat ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Low Molecular Weight ◽  
Wheat Cultivars ◽  
Glutenin Subunits ◽  
Molecular Weights ◽  
Additional Protein ◽  
Hmw Glutenin

Sodium dodecyl sulphate gradient polyacrylamide gel electrophoresis (SDSGPAGE) was used to resolve gliadin and high- and low-molecular-weight glutenin subunits from 19 registered Canadian spring wheat cultivars eligible for Canada Western Red Spring (CWRS) and Canada Prairie Spring (CPS) wheat grades and eight nonregistered spring wheat cultivars from the U.S.A. Reproducible molecular weight estimates were obtained for wheat proteins of apparent molecular weights ranging from 34 238 to 136 174 (avg. CV = 0.72%). Eight different patterns of HMW glutenin subunits consisting of 7–11 protein bands were observed for the 27 cultivars and their biotypes. SDSGPAGE was able to discriminate among the majority of cultivars with all non-registered cultivars and their biotypes distinguishable from registered cultivars. Separation of glutenin subunits along with gliadins provided additional protein bands which assisted in the discrimination of cultivars.Key words: SDS gradient PAGE, wheat cultivar identification, gliadin, glutenin subunits

Variability of Low-Molecular-Weight, Heat-Modifiable Outer Membrane Proteins of Neisseria meningitidis

1980 ◽  
Vol 30 (3) ◽  
pp. 642-648
Author(s):  
J. T. Poolman ◽  
S. De Marie ◽  
H. C. Zanen
Keyword(s):  
Molecular Weight ◽  
Outer Membrane ◽  
High Molecular Weight ◽  
Outer Membrane Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Low Molecular Weight ◽  
Molecular Weights ◽  
Sds Page ◽  
Index Cases

Analysis of major outer membrane protein (MOMP) profiles of various meningococci by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (SDS-PAGE) revealed the presence of 0 to 2 low-molecular-weight, heat-modifiable MOMPs (molecular weight, 25,000 to 32,000) and 1 to 3 high-molecular-weight MOMPs (molecular weight, 32,000 to 46,000). Heat modifiability was investigated by comparing MOMP profiles after heating in SDS solutions at 100°C for 5 min or at 40°C for 1 h. Low-molecular-weight MOMPs shifted to higher apparent molecular weights after being heated at 100°C. Heat modifiability of high-molecular-weight MOMPs varied among strains; whenever modified these proteins shifted to lower apparent molecular weights after complete denaturation. Variability of low-molecular-weight, heat-modifiable MOMPs was demonstrated when MOMP profiles were compared of (i) isolates from index cases and associated cases and carriers among contacts, (ii) different isolates from the same individual, and (iii) isolates from a small epidemic caused by serogroup W-135. In some cases high-molecular-weight MOMPs revealed quantitative differences among related strains. The observed variability and quantitative differences indicate that MOMP serotyping and typing on the basis of SDS-PAGE profiles (PAGE typing) need careful reevaluation.

Heterozygous Abnormal Fibrinogen Osaka III with the Replacement of γ Arginine-275 by Histidine Has an Apparently Higher Molecular Weight γ-Chain Variant

1992 ◽  
Vol 68 (05) ◽  
pp. 534-538 ◽  
Author(s):  
Nobuhiko Yoshida ◽  
Shingi Imaoka ◽  
Hajime Hirata ◽  
Michio Matsuda ◽  
Shinji Asakura
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Tetraacetic Acid ◽  
Fibrin Monomer ◽  
Sds Page ◽  
Thrombotic Tendency ◽  
Chain Variant

SummaryCongenitally abnormal fibrinogen Osaka III with the replacement of γ Arg-275 by His was found in a 38-year-old female with no bleeding or thrombotic tendency. Release of fibrinopeptide(s) by thrombin or reptilase was normal, but her thrombin or reptilase time in the absence of calcium was markedly prolonged and the polymerization of preformed fibrin monomer which was prepared by the treatment of fibrinogen with thrombin or reptilase was also markedly defective. Propositus' fibrinogen had normal crosslinking abilities of α- and γ-chains. Analysis of fibrinogen chains on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the system of Laemmli only revealed the presence of abnormal γ-chain with an apparently higher molecular weight, the presence of which was more clearly detected with SDS-PAGE of fibrin monomer obtained by thrombin treatment. Purified fragment D1 of fibrinogen Osaka III also seemed to contain an apparently higher molecular weight fragment D1 γ remnant on Laemmli gels, which was digested faster than the normal control by plasmin in the presence of [ethy-lenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA).

scholarly journals Isolation and Characterization of Two Proteins fromMoraxella catarrhalis That Bear a Common Epitope

1998 ◽  
Vol 66 (9) ◽  
pp. 4374-4381 ◽  
Author(s):  
John C. McMichael ◽  
Michael J. Fiske ◽  
Ross A. Fredenburg ◽  
Deb N. Chakravarti ◽  
Karl R. VanDerMeid ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Bacterial Attachment ◽  
Vaccine Candidates ◽  
Isolation And Characterization ◽  
Sds Page

ABSTRACT The UspA1 and UspA2 proteins of Moraxella catarrhalisare potential vaccine candidates for preventing disease caused by this organism. We have characterized both proteins and evaluated their vaccine potential using both in vitro and in vivo assays. Both proteins were purified from the O35E isolate by Triton X-100 extraction, followed by ion-exchange and hydroxyapatite chromatography. Analysis of the sequences of internal peptides, prepared by enzymatic and chemical cleavage of the proteins, revealed that UspA1 and UspA2 exhibited distinct structural differences but shared a common sequence including an epitope recognized by the monoclonal antibody 17C7. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), purified UspA1 exhibited a molecular weight of approximately 350,000 when unheated and a molecular weight of 100,000 after being heated for 10 min at 100°C. In contrast, purified UspA2 exhibited an apparent molecular weight of 240,000 by SDS-PAGE that did not change with the length of time of heating. Their sizes as determined by gel filtration were 1,150,000 and 830,000 for UspA1 and UspA2, respectively. Preliminary results indicate the proteins have separate functions in bacterial pathogenesis. Purified UspA1 was found to bind HEp-2 cells, and sera against UspA1, but not against UspA2, blocked binding of the O35E isolate to the HEp-2 cells. UspA1 also bound fibronectin and appears to have a role in bacterial attachment. Purified UspA2, however, did not bind fibronectin but had an affinity for vitronectin. Both proteins elicited bactericidal antibodies in mice to homologous and heterologous disease isolates. Finally, mice immunized with each of the proteins, followed by pulmonary challenge with either the homologous or a heterologous isolate, cleared the bacteria more rapidly than mock-immunized mice. These results suggest that UspA1 and UspA2 serve different virulence functions and that both are promising vaccine candidates.

scholarly journals The interrelations between high- and low-molecular-weight forms of normal and mutant (Krabbe-disease) galactocerebrosidase

1980 ◽  
Vol 189 (1) ◽  
pp. 9-15 ◽  
Author(s):  
Yoav Ben-Yoseph ◽  
Melinda Hungerford ◽  
Henry L. Nadler
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Active Form ◽  
Sodium Dodecyl ◽  
Krabbe Disease ◽  
Low Molecular Weight ◽  
Molecular Weight Form

Galactocerebrosidase (β-d-galactosyl-N-acylsphingosine galactohydrolase; EC 3.2.1.46) activity of brain and liver preparations from normal individuals and patients with Krabbe disease (globoid-cell leukodystrophy) have been separated by gel filtration into four different molecular-weight forms. The apparent mol.wts. were 760000±34000 and 121000±10000 for the high- and low-molecular-weight forms (peaks I and IV respectively) and 499000±22000 (mean±s.d.) and 256000±12000 for the intermediate forms (peaks II and III respectively). On examination by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, the high- and low-molecular-weight forms revealed a single protein band with a similar mobility corresponding to a mol.wt. of about 125000. Antigenic identity was demonstrated between the various molecular-weight forms of the normal and the mutant galactocerebrosidases by using antisera against either the high- or the low-molecular-weight enzymes. The high-molecular-weight form of galactocerebrosidase was found to possess higher specific activity toward natural substrates when compared with the low-molecular-weight form. It is suggested that the high-molecular-weight enzyme is the active form in vivo and an aggregation process that proceeds from a monomer (mol.wt. approx. 125000) to a dimer (mol.wt. approx. 250000) and from the dimer to either a tetramer (mol.wt. approx. 500000) or a hexamer (mol.wt. approx. 750000) takes place in normal as well as in Krabbe-disease tissues.

scholarly journals Evaluation of winter wheat collection in terms of HMW- and LMW-glutenin subunits

2010 ◽  
Vol 46 (Special Issue) ◽  
pp. S96-S99 ◽  
Author(s):  
J. Bradová ◽  
L. Štočková
Keyword(s):  
Molecular Weight ◽  
Winter Wheat ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Low Molecular Weight ◽  
Glutenin Subunits ◽  
Bread Making ◽  
Wheat Varieties ◽  
Lmw Glutenin Subunits ◽  
Winter Wheat Varieties

The composition of high molecular weight (HMW-GS) and low molecular weight (LMW-GS) glutenin subunits was examined in a collection of 86 Czech registered winter wheat varieties. These proteins were analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis. An inter-varietal polymorphism of the HMW and LMW glutenin subunits was detected. Twenty-one different patterns for HMW were identified, and eighteen for the LMW-glutenins. The different alleles encoded at the six glutenin loci were determined. Three, six, and four alleles were observed, respectively at the <I>Glu-A1, Glu-</I>B1, and <I>Glu-D1 </I>loci (encoding high HMW-GS). Three, eight, and three alleles of LMW-GS were found, respectively, at the <I>Glu-A3, Glu- B3</I>, and <I>Glu-D3 </I>loci. The evaluated varieties were split into four categories of baking quality, and these variety groups were analyzed for the presence of different HMW-GS and LMW-GS alleles. While the alleles <I>Glu-B1c </I>(7+9), and <I>Glu-D1d </I>(5+10) were detected exclusively in bread wheat varieties, the alleles <I>Glu-B1d </I>(6+8), <I>Glu-D1a </I>(2+12), and <I>Glu-A3e/f </I>only occurred in those varieties that are not suitable for bread-making. &nbsp;

Catabolism of streptokinase and polyethylene glycol-streptokinase: evidence for transport of intact forms through the biliary system in the mouse

Blood ◽  
1990 ◽  
Vol 76 (1) ◽  
pp. 73-79 ◽  
Author(s):  
FH Brucato ◽  
SV Pizzo
Keyword(s):  
Molecular Weight ◽  
Polyethylene Glycol ◽  
Gel Electrophoresis ◽  
Proteinase Inhibitors ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gi Tract ◽  
Substantial Fraction ◽  
Sds Page ◽  
Derivatives Of

Abstract The catabolism of streptokinase (SK) and polyethylene glycol derivatives of SK (PEG-SK) were studied in mice. The clearance and catabolism of SK:plasmin (SK:Pm) and PEG-SK:Pm activator complexes were also investigated. Native 125I-SK cleared rapidly (t1/2 = 15 minutes) from the circulation, with the majority of the ligand accumulating in the liver and gastrointestinal (GI) tract and a substantial fraction also localizing in the kidneys. SK, which was removed from the plasma by the liver, was secreted into bile and then the GI tract. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that 125I-SK recovered from liver and bile was homogeneous and of the same molecular weight (mol wt approximately 50,200) as native SK. PEG-125I-SK cleared slowly (t1/2 greater than 200 minutes), with more than 80% of the preparation localizing in liver and GI tract. The PEG-125I-SK secreted into the bile was also intact. The bile containing 125I-SK was incubated with stoichiometric amounts of plasminogen and electrophoresed under nondenaturing conditions. This study demonstrated that the secreted SK was able to form SK:Pg complexes. SDS-PAGE also showed activation of 125I-Pg that was incubated with recovered bile containing the SK. 125I-SK:Pm catabolism was also studied. In these experiments, the mol wt approximately 42,000 fragment obtained when SK is cleaved by plasmin was found in the bile. This fragment of 125I-SK was not recovered as part of a complex with plasmin, consistent with our previous observations that catabolism of SK:Pm involves transfer of the plasmin to plasma proteinase inhibitors while SK is catabolized independently. By contrast, when PEG-125I-SK:Pm was injected into mice, only intact PEG-125I-SK was found in the bile, consistent with our previous observations that the PEG derivatization blocks its degradation by plasmin.

scholarly journals Serological grouping ofTreponema hyodysenteriae

1990 ◽  
Vol 105 (1) ◽  
pp. 79-85 ◽  
Author(s):  
D. J. Hampson ◽  
J. R. L. Mhoma ◽  
B. G. Combs ◽  
J. I. Lee
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Vaccine Development ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cross Reactivity ◽  
Low Molecular Weight ◽  
Serological Response ◽  
Double Immunodiffusion ◽  
Sds Page ◽  
Treponema Hyodysenteriae

SUMMARYTwo Australian isolates ofTreponema hyodysenteriaewhich did not fit within the current serological grouping system for these bacteria wrere examined by agarose gel double immunodiffusion tests (AGDP). Isolate Vic1 was serologically unique, and we propose that it becomes the type organism for a new sixth serological group ofT. hyodysenteriae(Group F). Isolate Q1 was unusual in that lipopolysaccharide (LPS) extracted from it reacted strongly in AGDP with serum raised against the type organism for serogroup D (A1), and also weakly with serum raised against the type organism for serogroup B (WA1). The nature of this cross-reactivity was examined by using cross-absorbed antisera in AGDP, and by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis.The pattern of serological cross-reactivity between Q1, A1 and WA1 was complex and was not fully defined, but the isolate Q1 apparently shared low molecular weight ‘serogroup’ LPS antigens with A1, and shared higher molecular weight LPS antigens with WA1. On this basis Q1 was designated as belonging to serogroup D, although it was recommended that this be qualified as D (B) to indicate the presence of weak cross-reactivity with serogroup B. Such serological cross-reactivity may have significance in relation to the development of immunity toT. hyodysenteriae. Isolate Q1 may be a potentially useful organism for vaccine development because of its ability to induce a good serological response to LPS of treponemes from both serogroups D and B.

scholarly journals Incidental Detection of Dent-2 Disease in an Infant with Febrile Proteinuria

2018 ◽  
Vol 27 (4) ◽  
pp. 392-395 ◽  
Author(s):  
Shpetim Salihu ◽  
Katerina Tosheska ◽  
Svetlana Cekovska ◽  
Velibor Tasic
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulfate ◽  
Clinical Presentation ◽  
Viral Infections ◽  
Sodium Dodecyl ◽  
Febrile Illness ◽  
Low Molecular Weight ◽  
Acute Febrile Illness ◽  
Low Molecular Weight Proteinuria ◽  
Sds Page

Objective: Febrile proteinuria is functional proteinuria and is seen as a transitory phenomenon during acute febrile illness, mainly viral infections. It is a benign phenomenon and clears promptly with resolution of the infection. Clinical Presentation and Intervention: In this report, we present a patient who was thought to have febrile proteinuria. Persistence of significant proteinuria after resolution of the infection prompted biochemical and genetic workup which led to the diagnosis of Dent-2 disease. Conclusion: We recommend the use of SDS-PAGE (sodium dodecyl sulfate electropheresis) for the detection of low molecular weight proteinuria.

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