Escherichia coli in vitro recombinant system transports FhaB mutants

• Published in: Ji chu yi xue yu lin chuang = Jichu yixue yu linchuang = Basic medical sciences and clinics Vol. 42; no. 4; pp. 565 - 569
• Main Author: HUANG Pi-ying, CAI Kun, WEI Liang-wan, WANG Xing-yuan, WANG Zhe, CHU Yin-di, FAN En-guo
• Format: Journal Article
• Language: Chinese
• Published: Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College 01.04.2022
• Subjects: two-partner secretion|mutants|fhac|fhab
• ISSN: 1001-6325
• DOI: 10.16352/j.issn.1001-6325.2022.04.003

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.