RESULTS AND DISCUSSION
Initial engineering of C. ljungdahlii for butyrate production: strain B1.
Enhancing Crt expression through ribosome binding site modification: strain B2.
Integration of butyrate pathway genes into chromosome: strain B3.
Inactivating Pta-dependent acetate synthesis: strain B4.
Inactivating Pta-dependent acetate synthesis and AdhE1-dependent ethanol synthesis: strain B5.
Inactivating Pta-dependent acetate synthesis and CoA transferase: strain B6.
MATERIALS AND METHODS
Strains and growth conditions.
Construction of butyrate pathway genes in plasmids (strains B1 and B2).
Integration of butyrate pathway genes on chromosome (strain B3).
Construction of Cre-lox system for C. ljungdahlii.
Inactivation of pta gene and integration of butyrate pathway genes (strain B4).
Inactivation of adhE1 gene (strain B5).
Inactivation of ctf gene (strain B6).
Reverse transcription-quantitative PCR (qRT-PCR).
Western blot analysis.
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