Establishing Of The Two-dimensional Electrophoresis (2DE) Map Database And Locating Of Marked Proteins In The Protoscolex Phase Of Echinococcus Granulosus

Objective: To establish the datebase of two-dimensional electrophoretogram of protoscolex. To prepare antisera with three recombined antigens which were obtained from protoscolex in our early work. Then to purify the specific antibody to identify the foregone protein dot which is marked on the two-dimensional electrophoretogram. To lay the fundament for the applications of two-dimensional electrophoretogram of protoscolex.Methods 1 sample preparation: We chose protoscolex as sample. 2 protein separation: We separated the protein by two-dimensional electrophoresis separating technique. 3 pictures collection, preservation and analysis: We collected pictures by the Scanner GS-800, analyzed the obtained two-dimensional electrophoretogram of protoscolex with the PDquest software, then we initially got the protein quantity of protoscolex and the data of isoelectric points and molecular weight of the proteins. 4 expression and purification: We prepared antisera by immunizing the New-Zealand rarebits with three recombination proteins of protoscolex. After purifying the antibodies and analyzing by western blot, we identified the foregone proteins as markers in two-dimensional electrophoretogram of protoscolex.Results In two-dimensional electrophoretogram of protoscolex, 240 protein dots were separated respectively. We have initially acquired the protein information of isoelectric points and molecular weight with software. There were three recombination proteins of Echinococcus granulosus, zw-5, 14-3-3, p-29 purified successfully. We have prepared the antibodies by the method of immunology, and accomplished the identification the three recombination proteins, zw-5, 14-3-3, p-29, to two-dimensional electrophoretogram of protoscolex.Conclusion 1 Two-dimensional electrophoretogram of Echinococcus granulosus has been built up initially. 2 We have got 240 protein dots and all the molecular weight and isoelectric points have been acquired by software on the base of two-dimensional electrophoretogram. 3 We have prepared the antibodies of recombination proteins of protoscolex, and we have also marked three foregone protein dots on the two-dimensional electrophoresis of protoscolex.