A T-extended vector using a green fluorescent protein as an indicator

T-extended vector (T-vector) is a useful tool for cloning PCR products dire ctly. We exploited a novel T-vector using a green fluorescent protein (GFP) as an indicator based on insertional inactivation. The brightest GFP mutan t was used for easy detection even under daylight. The 100 bp and 0.9 kb of PCR products were cloned, and the transformant colonies with inserts were adjudged by the fluorescent green-white screening. The GFP system was more sensitive to insertional inactivation than the P-galactosidase system at th e conventional insertion sites. (C) 2000 Published by Elsevier Science B.V. All rights reserved.