ISOLATION, SEQUENCING AND EXPRESSION OF THE GENE ENCODING A MAJOR PROTEIN FROM THE BACKTERIOPHAGE ASSOCIATED WITH BARTONELLA-HENSELAE

The gene encoding a 31-kDa major protein (Pap31) associated with the b acteriophage harbored in Bartonella henselae was cloned and sequenced. Analysis of the resulting sequence revealed an open reading frame of 837 nucleotides coding for a protein of 279 amino acids. pap31 was the n subcloned downstream of the lacZ promoter in pUC19. pap31 was amplif ied by polymerase chain reaction, and the linear amplicon was used as template for in-vitro transcription and translation. A protein with an apparent molecular mass of approximately 31 kDa was synthesized from this reaction. Upon analysis of the deduced aa sequence, a potential s ignal sequence and a consensus signal peptidase cleavage site were ide ntified, indicative that Pap31 is modified posttranslationally, and th e mature protein may be targeted to the host membrane. (C) 1997 Elsevi er Science B.V.