May
Preliminary idea determination
Search and screening of cholesterol degrading genes
Search for a suitable promoter: oleic acid inducer was finally identified
The design of gene combination
June
Gene synthesis and plasmid construction
July
Prepared the competent
Bacterial culture and plasmid extraction
Transformed the target gene into the chassis organism Escherichia coli Nissel 1917
Performed cell counting to detect bacterial growth status
August
Functional verification of IsmA and BCoAT genes, including:
Cholesterol was measured by phthalaldehyde
SCFAs production was measured by gas chromatography
The bacteria were lysed by sonication for BSH crude enzyme solution extraction
September
Qualitative detection of bile saline hydrolase
Prepared egg yolk medium and perform the detection of cholesterol degradation from food sources
Detection of BSH enzyme activity
October
Verified the function of oleic acid inducer by microplate reader and mRFP
Confirmed the expression of IsmA and BSH by SDS-PAGE
Determined the downstream products of IsmA conversion to cholesterol by UHPLC
Preparation and functional verification of ACA microcapsules
Others
Two: the experiment of the record scanning
1、Share documents online:
2. Scanning of laboratory record book:
