Construction method of monascus genetically engineered microorganism having high-yield product Monacolin K and application of monascus genetically engineered microorganism
A technology of genetically engineered bacteria and construction methods, which is applied in the field of microbial genetic engineering, can solve the problem of low yield of MonacolinK, and achieve the effect of convenient method and strong ability
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[0051] Example 1. Construction of MonacolinK-producing MonacolinK genetically engineered bacteria
[0052] 1. Overexpression plasmid construction technical route
[0053] The MonacolinK genetically engineered bacteria with high production of MonacolinK of the present invention is obtained by using pBC-Hygro plasmid as an overexpression plasmid and overexpressing mokC or mokD or mokE or mokI genes. The route is: extract Monascus RNA, reverse transcription into cDNA, and use cDNA as a template to amplify the target fragment. After cutting and ligating the target fragments separately, construct a recombinant plasmid. Taking pBC-hygro-mokC as an example, the technical route is as follows figure 1 Shown.
[0054] 2. Extraction of Monascus RNA
[0055] The transcriptome RNA is extracted after purification of the mycelium of Monascus purple M1 that has been fermented to the 8th day. The extraction method is a conventional method in the art. Take 7μL of RNA sample for 1% agarose gel electro...
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