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Fig. 4 | Microbial Cell Factories

Fig. 4

From: Multiplex genetic manipulations in Clostridium butyricum and Clostridium sporogenes to secrete recombinant antigen proteins for oral-spore vaccination

Fig. 4

Procedure for genetic manipulations in clostridia for heterologous protein secretion. Briefly, the genes of interest, promoters, and signal peptides were ligated into E. coli-Clostridium shuttle vectors. The AUU start codon in the signal peptide would be used when difficulties (SNPs or ISs) were observed in E. coli. Following the transformation into clostridia, Western blotting analysis was conducted to identify the ideal expression cassette at the same growth phase. Finally, the expression cassette was integrated into the Clostridium chromosome by the CRISPR-Cas12a system and further validated through Western blotting analysis

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Microbial Cell Factories

ISSN: 1475-2859

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