Microchip electrophoretic analysis of Phaseolin patterns and Its comparison with currently used SDS-PAGE techniques

Silva, Emanuel Marques da; Santos, Teresa Maria Marques dos; Ganança, José Filipe Teixeira; Slaski, Jan Jacek; Carvalho, Miguel Â. A. Pinheiro de

http://hdl.handle.net/10400.13/3167

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Authors

Silva, Emanuel Marques da

Santos, Teresa Maria Marques dos

Ganança, José Filipe Teixeira

Slaski, Jan Jacek

Carvalho, Miguel Â. A. Pinheiro de

Abstract(s)

The goal of this work was to compare reproducibility of phaseolin patterns of common bean obtained by two electrophoretic protein separation techniques including the conventional SDS-PAGE and an automated chip electrophoresis system. Five standard cultivars of common bean provided by the United States Department of Agriculture (Beltsville, Maryland) that represented five phaseolin types, T (Tendergreen), C (Contender) and S (Sanilac), B (Boyaca) and P (Pampa), were used in this study. Comparison of the phaseolin patterns revealed that the chip-on-a-lab electrophoresis provided a good reproducibility. The phaseolin polymorphism included four to seven polypeptides typical for the pattern composition of the T, C and S types. The polymorphism of the B and P patterns was also established. Phaseolin polypeptides separated by the microchip electrophoresis exhibited differences with respect to the molecular weights and electrophoretic mobility as compared to the SDS-PAGE technique. This phenomenon could be attributed to the absence of a solid separation phase in the microchip electrophoresis. Moreover, this technique has potential to substantially accelerate screening of large bean germplasm collections since it allows for the accurate analysis of the higher number of individual plants within accessions than the conventional, tedious and time consuming SDS-PAGE method.

Keywords

Microchip electrophoresis SDS-PAGE Technique validation Protein pattern Phaseolin . Faculdade de Ciências da Vida

URI

http://hdl.handle.net/10400.13/3167

Citation

Silva, E. M., Santos, T. M. M., Ganança, J. F. T., Slaski, J. J., & Carvalho, M. Â. P. (2013). Microchip electrophoretic analysis of phaseolin patterns and its comparison with currently used SDS-PAGE techniques. Chromatographia, 76(17-18), 1163-1169. DOI 10.1007/s10337-013-2511-x