Antibody Preparation and Protein-Protein interaction between XpsG and XpsI in Xanthomonas campestris

• Main Authors: Lee, Meng-Shiunn, 李孟訓
• Other Authors: Ling-Yan Chen, Te-Chung Liu
• Format: Others
• Language: zh-TW
• Online Access: http://ndltd.ncl.edu.tw/handle/54645368039999282513

碩士 === 中山醫學院 === 生物化學研究所 === 85 === XpsG,H,I and J,having homologous leader sequence of type IV pilin,arecytoplasmic membrane proteins that require for the secretion of extracellularproteins in Xanthomonas campestris pv. campestris. The objective of this researchis to study the protein-protein interaction between XpsG,H,I and J and the putativepilin-like structure. The specific aims include the construction of expression vector,the overexpression of recombinant proteins,the induction of antibody and investigationof the protein-protein interaction in XC1701. In this study,four expression vectors wereconstructed. Among them, the recombinant protein of XpsG and XpsI were expressed and purifiedby both pET32a and pQE9 expression vectors from E.coli BL21(DE3) and M15[pREP4] strain. Theantibodies against XpsG and XpsI were also obtained by immunizing rabbits with purified recombinantproteins. Gel filtration chromatography and Western blot analysis were then performed to analyze thecofraction of XpsD,XpsG and XpsI proteins. The preliminary data showed that XpsG and XpsI,which wereeluted in the same fraction,were not cofractionation with XpsD. The protein-protein interaction betweenXpsG and XpsI in XC1701 still need other experiment such as immunoaffinity chromatography analysis to bedirect evidence for perfomed.