E-Book Overview
Barry W. Hicks and a team of well-practiced experimentalists demonstrate the power and the versatility of the green fluorescent protein (GFP) marker with a cutting-edge collection of readily reproducible GFP techniques. These include methods for detecting and imaging GFP (FRET, BRET, and whole-body imaging); for using GFP to create and select for transgenic organisms, for its use as a biosensor; and for studying viruses. A valuable companion CD-ROM displays many color figures and videos from over half the chapters.
E-Book Content
Methods in Molecular Biology TM VOLUME 183 Green Fluorescent Protein Applications and Protocols Edited by Barry W. Hicks Bonus CD-ROM Included HUMANA PRESS Amplification of cDNA from Invertebrates 3 1 Amplification of Representative cDNA Samples from Microscopic Amounts of Invertebrate Tissue to Search for New Genes Mikhail V. Matz 1. Introduction Recently, we cloned six new green fluorescent protein (GFP)-like fluorescent proteins from five species of Antozoa (1), including one red-emitting variant (DsRed), which is now commercially available. This project did not require expeditions and collection of animals on reefs: In all cases, the starting material was just several milligrams of tissue (e.g., a tentacle tip of a sea anemone), collected from a specimen in a private aquarium. This truly noninvasive kind of study was possible because the approach of total cDNA amplification, which is extensively applied to various tasks and biological models in our lab. This chapter outlines several year’s of experience in this helpful technique. The possibility of amplifying total cDNA obtained from small amounts of biological material is not yet routinely considered, despite the fact that obtaining amounts of material suitable for direct processing by standard methods is often ti