Prokaryotic Expression of Atp2 Protein of Rhodopseudomonas palustris and Preliminary Screening of Interaction Protein in Magnaporthe oryzae

doi:10.16409/j.cnki.2095-039x.2020.03.019

Abstract

Abstract: The earlier field experiment showed that PSB06 (Rhodopseudomonas palustris PSB-06) had a good inhibition effect on rice blast. In order to further exploit the biocontrol products, an ATP synthetic subunit Atp2 protein was identified from PSB06 culture medium by protein Chromatography., with the coding gene sequence of 1431 bp. ATP2 genes were cloned from photosynthetic bacteria PSB06, and the recombinant plasmid pET32a(+)-ATP2 was constructed. The recombinant Atp2 was highly purified by Affinity Chromatography and analyzed by SDS-PAGE and Western blot. Atp2 protein could significantly inhibit the appressorium formation of Magnaporthe oryzae and the virulence on the rice host. In order to elucidate its antibacterial mechanism, yeast-Two-Hybrid-System was employed with Atp2 as a bait protein to screen the proteins interacting with Atp2 in the cDNA library of M. oryzae. Seven positive proteins were obtained, including E3 ubiquitin ligase complex (MGG_04978), two ribosomal proteins (MGG_16204, MGG_04977) and four hypothetical proteins (MGG_09168, MGG_01034, MGG_03543, MGG_00718). The results provide theoretical basis for revealing the inhibition mechanism of Atp2 protein against M. oryzae.

Key words: Atp2, antibacterial mechanism, yeast two hybrid, protein interaction

CLC Number:

S476

WU Xiyang, LUO Luyun, TAN Xinqiu, ZHANG Deyong, CHEN Yue, LIU Yong. Prokaryotic Expression of Atp2 Protein of Rhodopseudomonas palustris and Preliminary Screening of Interaction Protein in Magnaporthe oryzae[J]. Chinese Journal Of Biological Control, 2020, 36(3): 421-428.

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