Virulent And Avirulent Comparative Proteomics Of Streptococcus Suis 2

To investigate Streptococcus suis 2 by protemeic technical about its virulence factors and pathogenic mechanism,we first get read for Streptococcus suis 2 sample using 2-DE,and to fiand the proper adding weight,the more important objective is fianding different proteion spots between highly virulent and avirulent.Fortunately,we found a way which adapt for two-dimensional gel electrophoresis(2-DE) to extract proteins of Streptococcus suis type2.In the process of samples preparation,we consider so many factors of interference.For example salt concentration,if salt concentration were so high , voltage may not reach the destination and proteins couldn't isoelectric condense completely,in the two-dimensional gel electrophoresis. In order to adapt to 2-DE, Gram-positive bacterium can be mixed by urea(high purification) and Tris(10mM/mL) in the process of samples preparation.Comparing A lysis buffer (urea and Tris)with B lysis buffer (urea and sulfocarbamid),we can conclude A lysis buffer is better B lysis buffer.In a word the lysis of urea is much better.And afer high speed centrifugation(low temperature), nucleic acid can be removed which is in the sample. In this way samples can run in the ampholine electrophoresis directly,and get a better result.Proteomics is a relatively newly coined term. It refers to the field of study of the proteome, which was defined for the first time in 1994 as"the PROTE in products of a genome".It used to consicider on resulotion and identify. Hence, as the method that offers the highest practical resolution in protein fractionation, two-dimensional gel electrophoresis (2-DE) is the most commonly used separation technique in proteomics. Its resolution power derives from orthogonally combining two separations based on two independent parameters: isoelectric focussing (IEF) separates based on charge and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) separates by size.We fiand a proper adding sample weight for 2-DE,in the end ,we receive a better gel map than fomer.Our result that a proper adding weight is about 150μg,A larger adding sample weight have a accumulative proteion in IEF,a smaller adding sample weight can't receive the siver staining compacity.Forther more our silver staining is not only the simplest metherd but also a best one . In the second dimension,we find Resolving gel buffer has a wrong pH,That is not only effect the pacing of gel but also the quality of gel .In this article proteomic techniques were used to study Streptococcus suis type 2.Whole cell proteins were extracted and performed 2-DE of different pH gradient.Comparaed to avirulent 1330 to highly virulent 449-1,6 proteion dots have different.one of 4 dots expression higher than avirulent,2 dots are interves. After different straines compared and identityfied, using ImageMaster 2D Platinum 6.0 analyses,fianding different proteion dots to the next identify.