Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot3945

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Screening Bacterial Colonies Using X-gal and IPTG: {alpha}-Complementation

Joseph Sambrook and David W. Russell

This protocol was adapted from Molecular Cloning, 3rd edition, by Joseph Sambrook and David W. Russell. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001

The first 15% of the full text of this article appears below.


INTRODUCTION

{alpha}-complementation occurs when two inactive fragments of E. coli ß-galactosidase associate to form a functional enzyme. Many plasmid vectors carry a short segment of DNA containing the coding information for the first 146 amino acids of ß-galactosidase. Vectors of this type are used in host cells that express the carboxy-terminal portion of the enzyme. Although neither the host nor the plasmid-encoded fragments of ß-galactosidase are themselves active, they can associate to form an enzymatically active protein. Lac+ . . . [Full Text of this Article]


MATERIALS


METHOD


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