The Analysis Of Protein Interaction Network Of Actinobacillus Pleuropneumoniae And Functional Validation Of Sub-networks

Proteins are the true implementation of the process of biological life and the basic of life activities. Protein interaction studies based on the life sciences is a important area, also is a window to understand the secret of life.Studies on the PPI is an important means to understand of biological processes, they plays an important role in functional genomics research. Porcine Contagious pleuropneumonia caused by Actinobacillus pleuropneumoniae (APP) is a respiratory disease of high contagious and mortality, causing severe economic losses globally in the breeding industry. The protein-protein interaction network of Actinobacillus pleuropneumoniae L20 was constructed by the method of homology protein mapping, we also predict and analysis the function of the overall network, and classify statistics of protions based on the degree, COG Categories. In addition, a bacteria two-hybrid assay was conducted in the network of HNS for functional verification.(1) Using the HPM method, a virtual protein interaction network of APP_L20 was constructed, consisting of 533 proteins and 2534 interaction pairs. By analysising of the total network of APP_L20 we can find out some new interaction relationship and mechanism of action, and even some new interacting pairs that are not easily verified or detected by experiments.(2) The analysis of APP sub-networks. There is a total of 19 hypothetical proteins involved in the network, which has taken the first step in analysising hypothetical proteins, it will be of great significance for revealing the function of hypothetical proteins. We summary the degrees of top 30 highly connected proteins in the network, and found that proteins of high degree are involved in the basic activities of bacterial cells organisms and the function of cells. Respectively we add up the COG of the whole genome and proteins in PPI network, we analysis the sub-networks of transcription, signal transduction, DNA replication recombination and repair to reveal its potential biological significance.(3) A bacterial two-hybrid assay was conducted.in the sub-network of HNS, and found 53.8% the contransformants grew well on the screening medium. Furtherly GST-pull down was used to verificate the positive interactions again. Actinobacillus pleuropneumoniae isolated from various places have a big difference in the sensitivity to antibiotics, due to long-term drug prevention and treatment it have a greater resistance, while the PPI analysising of APP_L20 can reveal potential signal channel, the ways of pathogenesis and discovery of new drug targets, it can offer a theoretical support for Actinobacillus pleuropneumoniae clinical prevention and treatment.