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Capillary electrophoresis separation of high molecular weight glutenin subunits in bread wheat (Triticum aestivum L.) and related species with phosphate-based buffers.
Electrophoresis. 2003 May; 24(9):1429-36.E

Abstract

This study focused on optimizing phosphate-based buffers and other capillary electrophoresis (CE) parameters for separating and characterizing high molecular weight glutenin subunits (HMW-GS) in bread wheat (Triticum aestivum L., AABBDD, 2n = 6x = 42), emmer (Triticum dicoccum, AABB, 2n = 4x = 28) and Aegilops tauschii (DD, 2n = 2x = 14). The fast and high-resolution separation of HMW-GS was achieved using 0.1 M phosphate-glycine buffer (pH 2.5, containing 20% acetonitrile and 0.05% hydroxypropylmethylcellulose) at 12.5 kV and 40 degrees C with 25 microm inside diameter (ID)x27 cm uncoated fused-silica capillary. In general, one sample separation can be analyzed in 15 min. The good run-to-run repeatable separation of HMW-GS could be obtained with a relative standard deviation of less than 1% when capillaries were rinsed with 1 M phosphoric acid for 2 min, followed by separation buffer for 2 min after each separation. The HMW-GS from some bread wheat cultivars as well as tetraploid and diploid accessions was separated by the CE method described above, and all subunits detected were well characterized and readily identified. Some HMW-GS showed reversed mobilities and elution order compared to the methods of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and SDS-CE. Particularly, most of the HMW-GS analyzed with the CE buffer used were separated into multiple peaks, generally a high peak plus a minor peak. CE appears to be capable of separating and characterizing HMW-GS with fast and high-resolution features, therefore it is expected to be useful for specific germplasm screening and desirable HMW-GS identification in wheat quality improvement.

Authors+Show Affiliations

Key Lab of Genetics and Biotechnology, Department of Biology, Capital Normal University, Beijing, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12731030

Citation

Yan, Yueming, et al. "Capillary Electrophoresis Separation of High Molecular Weight Glutenin Subunits in Bread Wheat (Triticum Aestivum L.) and Related Species With Phosphate-based Buffers." Electrophoresis, vol. 24, no. 9, 2003, pp. 1429-36.
Yan Y, Yu J, Jiang Y, et al. Capillary electrophoresis separation of high molecular weight glutenin subunits in bread wheat (Triticum aestivum L.) and related species with phosphate-based buffers. Electrophoresis. 2003;24(9):1429-36.
Yan, Y., Yu, J., Jiang, Y., Hu, Y., Cai, M., Hsam, S. L., & Zeller, F. J. (2003). Capillary electrophoresis separation of high molecular weight glutenin subunits in bread wheat (Triticum aestivum L.) and related species with phosphate-based buffers. Electrophoresis, 24(9), 1429-36.
Yan Y, et al. Capillary Electrophoresis Separation of High Molecular Weight Glutenin Subunits in Bread Wheat (Triticum Aestivum L.) and Related Species With Phosphate-based Buffers. Electrophoresis. 2003;24(9):1429-36. PubMed PMID: 12731030.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Capillary electrophoresis separation of high molecular weight glutenin subunits in bread wheat (Triticum aestivum L.) and related species with phosphate-based buffers. AU - Yan,Yueming, AU - Yu,Jianzhong, AU - Jiang,Yi, AU - Hu,Yingkao, AU - Cai,Minhua, AU - Hsam,Sai L K, AU - Zeller,Friedrich J, PY - 2003/5/6/pubmed PY - 2004/2/10/medline PY - 2003/5/6/entrez SP - 1429 EP - 36 JF - Electrophoresis JO - Electrophoresis VL - 24 IS - 9 N2 - This study focused on optimizing phosphate-based buffers and other capillary electrophoresis (CE) parameters for separating and characterizing high molecular weight glutenin subunits (HMW-GS) in bread wheat (Triticum aestivum L., AABBDD, 2n = 6x = 42), emmer (Triticum dicoccum, AABB, 2n = 4x = 28) and Aegilops tauschii (DD, 2n = 2x = 14). The fast and high-resolution separation of HMW-GS was achieved using 0.1 M phosphate-glycine buffer (pH 2.5, containing 20% acetonitrile and 0.05% hydroxypropylmethylcellulose) at 12.5 kV and 40 degrees C with 25 microm inside diameter (ID)x27 cm uncoated fused-silica capillary. In general, one sample separation can be analyzed in 15 min. The good run-to-run repeatable separation of HMW-GS could be obtained with a relative standard deviation of less than 1% when capillaries were rinsed with 1 M phosphoric acid for 2 min, followed by separation buffer for 2 min after each separation. The HMW-GS from some bread wheat cultivars as well as tetraploid and diploid accessions was separated by the CE method described above, and all subunits detected were well characterized and readily identified. Some HMW-GS showed reversed mobilities and elution order compared to the methods of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and SDS-CE. Particularly, most of the HMW-GS analyzed with the CE buffer used were separated into multiple peaks, generally a high peak plus a minor peak. CE appears to be capable of separating and characterizing HMW-GS with fast and high-resolution features, therefore it is expected to be useful for specific germplasm screening and desirable HMW-GS identification in wheat quality improvement. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/12731030/Capillary_electrophoresis_separation_of_high_molecular_weight_glutenin_subunits_in_bread_wheat__Triticum_aestivum_L___and_related_species_with_phosphate_based_buffers_ L2 - https://doi.org/10.1002/elps.200390184 DB - PRIME DP - Unbound Medicine ER -