Production of a monoclonal antibody specific for high molecular weight glutenin subunits (HMW-GS) in wheat and its antigenic determinant

2005 ◽  
Vol 48 (1) ◽  
pp. 89 ◽  
Author(s):  
Hanqian WANG
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibody ◽  
High Molecular Weight ◽  
Antigenic Determinant ◽  
Glutenin Subunits

An approach for isolating high-molecular-weight glutenin subunit genes using monoclonal antibodies

Genome ◽  
2006 ◽  
Vol 49 (2) ◽  
pp. 181-189 ◽  
Author(s):  
H Q Wang ◽  
X Y Zhang
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibody ◽  
Triticum Aestivum ◽  
Amino Acid ◽  
High Molecular Weight ◽  
Storage Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Glutenin Subunits ◽  
Sds Page

High-molecular-weight glutenin subunits (HMW-GSs) play an important role in the breadmaking quality of wheat flour. In China, cultivars such as Triticum aestivum 'Xiaoyan No. 6' carrying the 1Bx14 and 1By15 glutenin subunits usually have attributes that result in high-quality bread and noodles. HMW-GS 1Bx14 and 1By15 were isolated by preparative sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and used as an antigen to immunize BALB/c mice. A resulting monoclonal antibody belonging to the IgG1 subclass was shown to bind to all HMW-GSs of Triticum aestivum cultivars, but did not bind to other storage proteins of wheat seeds in a Western blot analysis. After screening a complementary DNA expression library from immature seeds of 'Xiaoyan No. 6' using the monoclonal antibody, the HMW-GS 1By15 gene was isolated and fully sequenced. The deduced amino acid sequence showed an extra stretch of 15 amino acid repeats consisting of a hexapeptide and a nonapeptide in the repetitive domain of this y-type HMW subunit. Bacterial expression of a modified 1By15 gene, in which the coding sequence for the signal peptide was removed and a BamHI site eliminated, gave rise to a protein with mobility identical to that of HMW-GSs extracted from seeds of 'Xiaoyan No. 6' via SDS-PAGE. This approach for isolating genes using specific monoclonal antibody against HMW-GS genes is a good alternative to the extensively used polymerase chain reaction (PCR) technology based on sequence homology of HMW-GSs in wheat and its relatives.Key words: wheat, HMW-GS, monoclonal antibody, immunoscreen.

scholarly journals Mitotic Illegitimate Recombination Is a Mechanism for Novel Changes in High-Molecular-Weight Glutenin Subunits in Wheat-Rye Hybrids

PLoS ONE ◽  
2011 ◽  
Vol 6 (8) ◽  
pp. e23511 ◽  
Author(s):  
Zhongwei Yuan ◽  
Dengcai Liu ◽  
Lianquan Zhang ◽  
Li Zhang ◽  
Wenjie Chen ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Illegitimate Recombination ◽  
Glutenin Subunits

scholarly journals Effect of High Molecular Weight Glutenin Subunits on Bread-Making Quality Using Near-Isogenic Lines.

2000 ◽  
Vol 50 (4) ◽  
pp. 303-308 ◽  
Author(s):  
Kanenori Tanaka ◽  
Hiroaki Yamauchi ◽  
Zenta Nishio ◽  
Tatsuo Kuwabara
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Near Isogenic Lines ◽  
Glutenin Subunits ◽  
Bread Making ◽  
Isogenic Lines

Simultaneous Isolation of Wheat High Molecular Weight and Low Molecular Weight Glutenin Subunits

1998 ◽  
Vol 28 (1) ◽  
pp. 25-32 ◽  
Author(s):  
I.M. Verbruggen ◽  
W.S. Veraverbeke ◽  
A. Vandamme ◽  
J.A. Delcour
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Low Molecular Weight ◽  
Glutenin Subunits
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