High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method

Abstract

Here we describe a high-efficiency version of the lithium acetate/single-stranded carrier DNA/PEG method of transformation of Saccharomyces cerevisiae. This method currently gives the highest efficiency and yield of transformants, although a faster protocol is available for small number of transformations. The procedure takes up to 1.5 h, depending on the length of heat shock, once the yeast culture has been grown. This method is useful for most transformation requirements.

Reconstitution of a 10-gene pathway for synthesis of the plant alkaloid dihydrosanguinarine in Saccharomyces cerevisiae

“…Transformations were either performed by heat shock in the presence of lithium acetate, carrier DNA and PEG 3350 (ref. 60), or by electroporation in the presence of sorbitol 9 .…”

Dual Function of the Cytochrome P450 CYP76 Family from Arabidopsis thaliana in the Metabolism of Monoterpenols and Phenylurea Herbicides

“…The WAT11 yeast strain was transformed with pYeDP60u2 containing the different P450 sequences as described in Gietz and Schiestl (2007). Yeast cultures were grown and P450 expression was induced as described in Pompon et al (1996).…”

Polymorphisms in the LAC12 gene explain lactose utilisation variability in Kluyveromyces marxianus strains

“…All yeast 24 transformations were performed using the standard Lithium Acetate 25 procedure (Gietz and Schiestl, 2007). Plasmids for functional expression of 1 LAC12 genes in S. cerevisiae were constructed using the pGREG-505 2 backbone (Jansen et al, 2005).…”