期刊
JOURNAL OF SEPARATION SCIENCE
卷 34, 期 19, 页码 2618-2630出版社
WILEY PERIODICALS, INC
DOI: 10.1002/jssc.201100238
关键词
Expression(E); Identity(E); NanoLC-MSE; ProteinLynx global server; Soybean
资金
- Brazilian Agricultural Research Corporation
- National Council for Scientific and Technological Development (CNPq)
- Fundacao de Apoio a Pesquisa-DF (FAP-DF)
The use of mass spectrometry to identify recombinant proteins that are expressed in total soluble proteins (TSPs) from plant extracts is necessary to accelerate further processing steps. For example, the method consists of TSP sample preparation and trypsin digestion prior to the preliminary characterization using nanoUPLC-MSE analysis of the recombinant proteins that are expressed in TSP samples of transgenic soybean seeds. A TSP sample as small as 50 mu g can be effectively analyzed. In this study, transgenic soybean seeds that expressed recombinant cancer testis antigen (CTAG) were used. The procedure covered 30% of the protein sequence and was quantified at 0.26 ng, which corresponded to 0.1% of the TSP sample. A comparative proteomic profile was generated by the comparison of a negative control and sample that showed a unique expression pattern of CTAG in a transgenic line. The experimental data from the TSP extraction, sample preparation and data analysis are discussed herein.
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