TRANSFORMATION OF PICHIA METHANOLICA

摘要

Methods for transforming Pichia methanolica, and DNA molecules useful in transformation of P. methanolica, are disclosed. P. methanolica cells are exposed, in the presence of DNA molecules, to a pulsed electric field having a field strength of from 2.5 to 4.5 kV/cm and a pulse duration of from 1 to 40 milliseconds, whereby the DNA molecules are introduced into the cells. The DNA molecules may comprise an expression unit that includes a transcription promoter of a P. methanolica gene operably linked to a segment encoding a polypeptide or protein of interest. The DNA molecules may also encode a selectable marker, such as a P. methanolica ADE2 gene. Cells transformed according to the invention may be used in production systems for the preparation of proteins of commercial importance.