Wyckoff, E., L. Sampson, M. Hayden, R. Parr, W. M. Huang, and S. Casjens. 1986. Plasmid vectors useful in the study of translation initiation signals. Gene 43:281-286.
The construction and characterization of plasmid vectors useful in the
analysis of translation initiation signals in Escherichia coli and in the
construction of lacZ gene hybrids are described. Transcription on the vectors
proceeds from a cAMP-independent lac promoter through several restriction sites
into a truncated lacZ structural gene lacking its first eight codons. Because
this gene has no translation initiation signal, its level of expression is
extremely low. A DNA fragment containing a translation start signal can be
inserted between the promoter and truncated lacZ gene to produce a hybrid
protein with functional beta-galactosidase activity. The vectors described here
differ in sequence between the EcoRI cloning site and the lacZ gene to allow
easy, in-frame joining of DNA containing a translation initiation signal to the
lacZ gene. Cells containing plasmids can be screened directly for in-frame
inserts by colony color on indicator plates.
