A positive screen for cloning PCR products.
We have developed a positive screen for cloning PCR products based on translational activation of lacZ. A vector with a translationally deficient lacZ alpha gene has been made by deletion of the Shine-Dalgarno sequence and initiation codon. The Shine-Dalgarno sequence and initiation codon are incorporated into one of the PCR primers to allow complementation by the PCR product of the inactive lacZ alpha gene, which results in blue transformed bacterial colonies. This screen allows more efficient detection of clones containing inserts made by PCR.
Keese
P
P
CSIRO, Division of Plant Industry, Canberra, Australia.
Graf
L
L
D003001:Cloning, Molecular / Q000379:methods*; D018387:Codon, Initiator; D005786:Gene Expression Regulation; D007763:Lac Operon; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D012641:Selection, Genetic
D018387:Codon, Initiator