A Study on the Interaction of Streptomyces Coelicolor Proteins (TpgC and TapC) Using a SPR Sensor

• Main Authors: Ying-Ren Lin, 林英仁
• Other Authors: Jui-Chuang Wu
• Format: Others
• Language: en_US
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/75775407160910913613

碩士 === 中原大學 === 化學工程研究所 === 97 === The genus Streptomyces is a gram-positive soil dwelling bacterium that currently produces over two-thirds of the naturally derived antibiotics in human and veterinary medicine. Streptomyces is unusual among bacteria for having protein-capped linear plasmids and chromosomes. The terminal protein gene (Tpg) is required for the specific replication of Streptomyces chromosomes or plasmids as linear molecules, and the telomere associated protein (Tap) is essential for the propagation of Streptomyces chromosomes and plasmids in linear form. In this work, a custom-built surface plasmon resonance (SPR) sensor based on wavelength interrogation with a four-channel Teflon flow cell was used to investigate the interaction between Tpg and Tap from the specie Streptomyces coelicolor, which is genetically the best known representative of the genus Streptomyces. The target proteins, herein called TpgC and TapC, were initially His-tagged and expressed in Escherichia coli (E. coli). The bacterial cultures were purified using immobilized metal ion affinity chromatography (IMAC) and dialyzed before running in the SPR sensor. A methyl-terminated self-assembled monolayer (SAM) immobilized on the sensor substrate was used in all SPR experiments. The amount of proteins immobilized on the sensor surface was computed based on the quantitative mathematical formalism established by Jung et al.