Methods for transforming yeast

摘要

This invention is directed to the transformation of yeast, and mutants thereof, by electroporation, which result in stably transformed yeast host cells that express recombinant products. This invention also is directed to transformed yeast cells and libraries.

主权项

1. A method of preparing a yeast library by electroporation of yeast cells comprising the steps of:
incubating the yeast cells in a solution comprising 0.01 to 1.0 M lithium acetate (LiAc) and 1 to 100 mM dithiothreitol (DTT), providing a suspension comprising vector DNA, insert DNA, the incubated yeast cells, 0.1 to 10 M sorbitol, and 0.1 to 10 mM CaCl2 or MgCl2, and electroporating the suspension at 5 kV/cm to 12.5 kV/cm with a capacitance of about 10 to about 50 μF; wherein the yeast cells are transformed with a transformation efficiency greater than 1×108 transformed yeast cells per μg of nucleic acid vector, thereby to obtain a library size of at least 1×109.