Jichu yixue yu linchuang (Apr 2022)

Escherichia coli in vitro recombinant system transports FhaB mutants

  • HUANG Pi-ying, CAI Kun, WEI Liang-wan, WANG Xing-yuan, WANG Zhe, CHU Yin-di, FAN En-guo

DOI
https://doi.org/10.16352/j.issn.1001-6325.2022.04.003
Journal volume & issue
Vol. 42, no. 4
pp. 565 – 569

Abstract

Read online

Objective To explore the mechanism of two-partner secretion with an in vitro reconstitution system. Methods Spheroplasts were used to construct FhaB mutants with different molecular weights, and then combined with FhaC containing proteoliposomes to perform the translocation experiments followed by digestion with proteinase K to test the transportation of substrates with different sizes in vitro by this recombinant system. Results After successful construction of the mutant, 37 ku, 60 ku and 80 ku bands secreted by spheroplasts were detected, and FhaC-containing protein liposomes could transport these three substrates with molecular weight as high as 80 ku. Conclusions The recombinant transport system constructed in vitro may transport substrates with large molecular weight and the mechanism of the two-partner secretion can be studied by this system.

Keywords