Thermotolerance improvement of engineered Saccharomyces cerevisiae ERG5 Delta ERG4 Delta ERG3 Delta, molecular mechanism, and its application in corn ethanol production

Table 1 Prepared primers for amplification of genes and plasmids

Primers	Sequences (5′ → 3′)	Sizes
ERG5-gRNA-F1	ATTTCATGGAAAAAGACCTGGGGGTTTTAGAGCTAGAAATAGCAAG	6509
ERG5-gRNA-R1	CCCCAGGTCTTTTTCCATGAAATGATCATTTATCTT TCACTGCGGA	bp
ERG4-gRNA-F1	CTATGTGACACCACAATTGGGGGGTTTTAGAGCTAGAAATAGCAAG	6509
ERG4-gRNA-R1	CCCCCAATTGTGGTGTCACATAGGATCATTTATCTTTCACTGCGGA	bp
ERG3-gRNA-F1	AAGATTGATTATGAAAACCACGGGTTTTAGAGCTAGAAATAGCAAG	6509
ERG3-gRNA-R1	CCGTGGTTTTCATAATCAATCTTGATCATTTATCTTTCACTGCGGA	bp
dDNA-MFC-F	GTTCCGTATCGCACACGCCG	628
dDNA-MFC-R	CTAGCTAACATTAATGTTGA	bp
dDNA-XYNA-F	CCCCACACACCATAGCTTCA	1129
dDNA-XYNA-R	GCGGATGTGGGGGGAGGGC	Bp
dDNA-CEL-F	CCCCACACACCATAGCTTCA	880
dDNA-CEL-R	CCGCCTGCGCCGCTCCGGTG	bp

ERG5-gRNA, ERG4-gRNA, and ERG3-gRNA primers were used to construct the vectors for ERG5-gRNA, ERG4-gRNA, and ERG3-gRNA, respectively. Three pairs of primers for the amplification of MFC (Ampullaria crossean multi-functional cellulase, 628 bp), XYNA (Endo-1,4-beta-xylanase, 1129 bp), and CEL (Cellulase, 880 bp) were used to knock-out ERG5, ERG4, and ERG3 as donor DNA, respectively

ISSN: 2731-3654