Unit

UNIT 13.9 Manipulation of Plasmids from Yeast Cells

  1. Victoria Lundblad1,
  2. Heng Zhou2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1309s39

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Lundblad, V. and Zhou, H. 2001. Manipulation of Plasmids from Yeast Cells. Current Protocols in Molecular Biology. 39:III:13.9:13.9.1–13.9.6.

Author Information

  1. 1

    Baylor College of Medicine, Houston, Texas

  2. 2

    Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUL 1997

Abstract

This unit describes several procedures for manipulating plasmids in yeast cells. The first is a general method to segregate autonomously replicating plasmids from cells: plasmid-containing yeast cells are grown in nonselective medium, and colonies lacking the plasmid are identified by replica plating. The second, plasmid shuffling, represents a specialized version of plasmid segregation that is useful for analyzing the function of essential genes and for identifying conditional lethal mutations in essential genes. The third approach, plasmid gap repair, is based on the efficient homologous recombination characteristics of yeast cells. Plasmid gap repair can be be used as a method to incorporate mutagenized DNA fragments into a yeast plasmid, rescue genomic mutations onto plasmids, or map alleles of a given gene.