Yeast YIp\|type expression recombinant plasmid(pCMA2\|1) was constructed. The expression of α\|acetolactate decarboxylase gene from \%Bacillus subtilis\% was controled by \%CUP1\% promoter and its own terminator. The...Yeast YIp\|type expression recombinant plasmid(pCMA2\|1) was constructed. The expression of α\|acetolactate decarboxylase gene from \%Bacillus subtilis\% was controled by \%CUP1\% promoter and its own terminator. The recombinant plasmid pCMA2\|1 was introduced into the brewer’s yeast PJ3\|5. Transformants were selected using copper resistance as selected marker. The results of activity assay showed that PJ3\|5 didn’t produce α\|acetolactate decarboxylase(ALDC), where as the activity of α\|ALDC in transformants were induced by copper sulfate. The laboratory scale fermentation test confirmed that the total diacetyl concentration was reduced effectively by α\|ALDC in transformant.展开更多
The partial genomic library of Saccharomyce cerevisiae FL189 possessing strong flocculation ability was constructed using Yeast-E.coli shuttle plasmid YCp50 as vector.Recombinant plasmid containing flocculation gene w...The partial genomic library of Saccharomyce cerevisiae FL189 possessing strong flocculation ability was constructed using Yeast-E.coli shuttle plasmid YCp50 as vector.Recombinant plasmid containing flocculation gene was obtained by screening the growth of transformants on the selective medium and measurement flocculation,designated as pCF1.pCF1 was introduced into industrial yeast strain PJ208-5-15.Six transformants PJ208-5-15-1(pCF1)~PJ208-5-15-6(pCF1)possessing strong flocculation ability were obtained.The results of Southern blot and restriction endonuclease analysis showed that the insert is about 4.3kb and could hybridize with the probe (2.6kb EcoRV fragment of FLO1).Flocculation ability assay indicated that the transformants possess strong flocculation ability.Hence,the gene controlling flocculation phenotype exists in the cloned DNA fragment.The restriction endonuclease analysis and the sequence analysis of the insert DNA fragment are in progress.展开更多
文摘Yeast YIp\|type expression recombinant plasmid(pCMA2\|1) was constructed. The expression of α\|acetolactate decarboxylase gene from \%Bacillus subtilis\% was controled by \%CUP1\% promoter and its own terminator. The recombinant plasmid pCMA2\|1 was introduced into the brewer’s yeast PJ3\|5. Transformants were selected using copper resistance as selected marker. The results of activity assay showed that PJ3\|5 didn’t produce α\|acetolactate decarboxylase(ALDC), where as the activity of α\|ALDC in transformants were induced by copper sulfate. The laboratory scale fermentation test confirmed that the total diacetyl concentration was reduced effectively by α\|ALDC in transformant.
文摘The partial genomic library of Saccharomyce cerevisiae FL189 possessing strong flocculation ability was constructed using Yeast-E.coli shuttle plasmid YCp50 as vector.Recombinant plasmid containing flocculation gene was obtained by screening the growth of transformants on the selective medium and measurement flocculation,designated as pCF1.pCF1 was introduced into industrial yeast strain PJ208-5-15.Six transformants PJ208-5-15-1(pCF1)~PJ208-5-15-6(pCF1)possessing strong flocculation ability were obtained.The results of Southern blot and restriction endonuclease analysis showed that the insert is about 4.3kb and could hybridize with the probe (2.6kb EcoRV fragment of FLO1).Flocculation ability assay indicated that the transformants possess strong flocculation ability.Hence,the gene controlling flocculation phenotype exists in the cloned DNA fragment.The restriction endonuclease analysis and the sequence analysis of the insert DNA fragment are in progress.