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. 2019 May 29;10:1177. doi: 10.3389/fmicb.2019.01177

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Copyright © 2019 Su, Jin, Wu, Fei, Li, Guo, Feng and Xiao.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Production and purification of recombinant MEDLE proteins CpMEDLE-3 and CpMEDLE-5 from Cryptosporidium parvum and ChMEDLE-1 from Cryptosporidium hominis. (A) PCR amplification of the target gene in genomic DNA. Lane M: molecular markers; lane 1: PCR product. (B) Expression of recombinant MEDLE protein in E. coli BL21 (DE3) as revealed by SDS-PAGE analysis. Lane M: molecular weight markers; lane 1: lysate from bacteria culture transformed with the recombinant plasmid without IPTG induction; lane 2: lysate from similar bacteria culture induced by IPTG for 2 h; lane 3: lysate from bacteria culture induced by IPTG for 8 h, with the expected product indicated by an arrow. (C) Western blot analysis of the recombinant protein. Lane M: molecular weight markers; lane 1: lysate from bacteria culture transformed with recombinant plasmid without IPTG induction; lane 2: supernatant from IPTG-induced bacterial culture; lane 3: cell lysate from IPTG-induced bacterial culture. (D) Purification of recombinant proteins. Lane M: molecular weight markers; lane 1: purified recombinant proteins from Ni-NTA affinity chromatography.