Isolation and Mapping of Small Cauliflower Mosaic Virus DNA Fragments Active as Promoters in Escherichia coli

Abstract

Small EcoRI(*) fragments of cauliflower mosiac virus DNA (strain CM4-184), which act as promoters for the tetracycline resistance gene on the promoter probe plasmid pBRH4 in Escherichia coli, have been isolated and mapped on the viral genome. Two regions of the viral genome contain DNA sequences with promoter activity in E. coli. Two independent cloned fragments from one region direct a high level of tetracycline resistance (up to 38 mug of tetracycline per ml). Two independent fragments from the second region of the viral genome also direct tetracycline resistance, but at lower levels. The activity of the two fragments with the strongest promoter activity in E. coli may direct transcription of the viral genome in a clockwise direction. This is consistent with the direction of transcription predicted from sequence analysis of the viral DNA (Franck et al., Cell 21: 285-294, 1980). One of these fragments maps at the start of a large open translational reading frame which is predicted to contain the coding sequence for the viral coat protein. Each promoter-active fragment is located in the 5'-terminal portion of one of the six open reading frames predicted from the DNA sequence.