Alpha complementation of LacZ in mammalian cells

Alpha complementation of LacZ in mammalian cells.

The bacterial lactose converting enzyme beta-galactosidase (β-gal) and its gene (LacZ) have been studied for many years (1 and references therein) and are among the most utilized tools in molecular biology. The LacZ product, a polypeptide of 1029 amino acids, gives rise to the functional enzyme after tetramerization (2 and references therein) and is easily detected by chromogenic substrates eit...

Gene expression and cell fusion analyzed by lacZ complementation in mammalian cells.

Complementing reporter genes provide biological indicators of coincident expression of proteins in cells. We have adapted intracistronic complementation of the Escherichia coli lacZ gene for use in mammalian cells. Enzymatic activity detectable by quantitative biochemical assay, flow cytometry, or microscopy is produced upon convergent expression of two distinct mutant lacZ peptides within sing...

Characterization of lacZ complementation deletions using membrane receptor dimerization.

We describe the screening of lacZ deletions in mammalian cells and the discovery of a novel pair of lacZ deletions that can undergo alpha-complementation only when they are fused to peptides that interact with each other. The two lacZ deletions, delta N 11-75 and delta C 82-1023, were first characterized by fusing to two small interacting peptides and were then further analyzed by fusing to thr...

Construction of an Escherichia-Pseudomonas shuttle vector containing an aminoglycoside phosphotransferase gene and a lacZ'' Gene for alpha-complementation.

A new 4.87 kb Escherichia-Pseudomonas shuttle vector has been constructed by inserting a 1.27 kb DNA fragment with a replication origin of a Pseudomonas plasmid pRO1614 into the 3.6 kb E. coli plasmid pBGS18. This vector, designated pJH1, contains an aminoglycoside phosphotransferase gene (aph) from Tn903, a lacZ'' gene for alpha-complementation and a versatile multiple cloning site possessing ...

Dual chromogenic reporter gene detection in mammalian cells with lacZ and arabinofuranosidase.