Expression of FPPS Recombinant Protein from Pogostemon cablin and Screening of the Interaction Proteins

doi:10.13560/j.cnki.biotech.bull.1985.2019-0475

Abstract

Abstract: The purpose of this study is to express the recombinant protein of farnesyl pyrophosphatase synthase（PatFPPS）of Pogostemon cablin in prokaryotic cells and to screen its interaction proteins. The PatFPPS CDS was amplified and ligated into the pGEX-6P-1 vector by PCR. The plasmid confirmed by sequencing was obtained and transformed into BL21（DE3）expression strain,which was then induced by IPTG and the GST-tagged PatFPPS fusion protein was acquired. With GST Pull-Down technique,GST-tagged PatFPPS fusion protein was incubated with total proteins from the P. cablin leaves in vitro. The solution containing the protein complex was eluted and then identified by SDS-PAGE and LC-MS/MS. The results showed that the prokaryotic expression system of PatFPPS was successfully established and the purified recombinant protein was obtained,and some candidate interaction proteins were screened. In conclusion,soluble PatFPPS protein can be obtained with optimized prokaryotic expression system,and candidate proteins interacted with PatFPPS are identified.

Key words: Pogostemon cablin, protein expression, patchouli alcohol, interaction protein, methyl jasmonate

ZHONG Li-ting, CHEN Xiu-zhen, TANG Yun, LI Jun-ren, WANG Xiao-bing, LIU Yan-ting, ZHOU Xuan-xuan, ZHAN Ruo-ting, CHEN Li-kai. Expression of FPPS Recombinant Protein from Pogostemon cablin and Screening of the Interaction Proteins[J]. Biotechnology Bulletin, 2019, 35(12): 10-15.

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