Method for rapid optimization of yeast cell plants
A yeast cell and yeast technology, applied in the field of rapid optimization of yeast cell factories, can solve problems such as low purity of beta-carotene, and achieve the effects of low cost, high efficiency and improved efficiency
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Embodiment 1
[0031] Example 1: Construction of recombinant Saccharomyces cerevisiae strains producing β-carotene
[0032] Taking the yeast strain carrying the synV chromosome as the starting strain, four genes for the synthesis of β-carotene from different sources were obtained from GeneBank: GGPP synthase gene crtE, phytoene synthase gene crtB, The phytoene dehydrogenase gene crtI and the lycopene cyclase gene crtY, meanwhile, in order to integrate the four genes into the chromosome of Saccharomyces cerevisiae for stable expression, a yeast endogenous assembly strategy was adopted. About 40 bp of homology arms were designed between the obtained above gene fragments, and at the same time, the genes on the left and right sides were designed to be homologous to the CAN site of Saccharomyces cerevisiae synV chromosome, with a length of about 40 bp. The above-mentioned genes were all codon-optimized by Saccharomyces cerevisiae and properly avoided common restriction enzyme sites, and were arti...
Embodiment 2
[0033] Example 2: Optimization of recombinant Saccharomyces cerevisiae strain yYW245 producing β-carotene
[0034] 1. Construction of diploid Saccharomyces cerevisiae strain
[0035] Saccharomyces cerevisiae yYW0245(MAT a) was hybridized with artificial Saccharomyces cerevisiae yYW0233(MATα) carrying chromosome number synIII, wherein the 3' end of each complete functional element on chromosome number synIII was inserted into the nucleotide sequence shown in SEQ ID NO:1 (Using de novo synthesis to insert the nucleotide sequence shown in SEQ ID NO:1 into the 3' end of the original chromosome functional element on a large scale), the specific steps for the construction of the diploid Saccharomyces cerevisiae strain are as follows:
[0036] After the two kinds of yeast plates were streaked and activated, they were inoculated into 5ml YPD (20g / L peptone, 10g / L yeast powder, 22g / L glucose monohydrate, 20g / L agar powder, pH 6.1) liquid medium at the same time, at 30°C After culturin...
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