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Nucleic Acids Research, 1988, Vol. 16, No. 13 5727-5740
© 1988

Articles

Site-directed mutagenesis in the DNA linking site of bacteriophage ø29 terminal protein: isolation and characterization of a Ser232->Thr mutant

Cristina Garmendia, Margarita Salas and José M. Hermoso

Centro de Biología Molecular (CSIC-UAM), Universidad Autónoma Canto Blanco, 28049 Madrid, Spain

Received May 12, 1988. Accepted June 6, 1988.

By site-directed mutagenesis we have changed the serine residue 232 of the ø29 terminal protein, involved in the covalent linkage to dAMP for the initiation of replication, into a threonine residue. The mutant terminal protein has been purified to homogeneity and shown to be inactive in the formation of the initiation complex; nevertheless, the mutant protein retains its ability to interact with the ø29 DNA polymerase and with the DNA. The results obtained indicate a high specificity in the linking site of the terminal protein.


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