Analysis of the Xanthomonas campestris pv. campestris membrane proteins related to the infection of lytic phage L7

• Main Authors: Shen-Yen Lin, 林珊瑩
• Other Authors: Shu-Fen Weng
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/57844935550304087839

碩士 === 國立中興大學 === 分子生物學研究所 === 92 === Comparing the membrane protein of different Xanthomonas isolates (Xc11, Xc17, and XcP20H) by two dimention electrophoresis (2D), the result show that Xanthomonas isolates membrane protein is not different. On the other hand, there is no difference between the Xc17 and XKG by membrane protein comparison. The AmpG of Xc17 expression level is too low to detect by two dimension electrophoresis . The experiment of comparison the membrane protein expression in different growth phase, the result reveals that membrane protein expression is not different. The most membrane protein isn’t able to express after the L7 infect XcP20H 30 minutes, the result reveal that L7 will lyse P20H at 30 minutes. From the 2D gels of phage (L7) resistance strain (B236) and phage sensitive strain (XcP20H), there is a membrane protein on B236 which expression level is less than XcP20H.The result of MALTI-TOF knows that this protein in an outer membrane protein P6 precursor (OmpP6), it’s pI 5.5, MW 20.4kDa . Unfortunately, we can’t get the ompP6 mutant by several ways, so the gene may be an essential gene for bacterial growth. From the Western blotting knows that the OmpP6 is an outer membrane protein and the OmpP6 folding is affected by lipopolysaccharide . Gram negative bacteria needs the FeⅢ transporter, the FepA is a outer membrane porin for ferric enterobactin. The PfepAM is a fepA gene mutant , the bacteriophage can’t infect it. There is no difference the XcP20H and PfepAM of growth curve, virulence, and extracellular enzyme secretion. From the result of phage infection reveals that the FepA may be a receptor or component of receptor for L7.