LacZ gene vector expression

Ligated plasmids should be checked for insertion of the desired fragment instead of just religation, most conveniently by blue-white screening. In vectors with the lacZ gene sequence, insertion at any restriction site of the MCS results in disruption of that sequence so that no /i-galactosidase is expressed and the synthetic X-gal substrate cannot form blue colonies. 

TABLE 8.2 Some Commercial Plasmid Vectors Expressing Full-Length lacZ Gene... 

Molsberger G., Schafer U., and Schafer M. 1988. A new set of lacZ fusion vectors, pUCPlac, for studying gene expression in Drosophila by P-mediated transformation. Gene 63 147-151. 

Andersen JK, Garber DA, Meaney CA, Breakefield XO (1992) Gene transfer into mammalian central nervous system using herpes virus vectors Extended expression of bacterial lacZ in neurons using the neuron-specific enolase promoter. Hum Gene Ther 3 487-499. 

Using a vector carrying a splice acceptor sequence placed upstream of a reporter gene (such as lacZ or GFP [green fluorescent protein]), different types of regulatory or gene sequences can be trapped (31-33). ES cell-chimeric embryos are stained for the histochemical marker to reveal expression domains of the tr jped elements (31-33). On the basis of the expression pattern information... 

Figure 6. Structure of /3-glucuronidase expression cassettes. TER is a 260 bp polyadenylation signal from the nopaline synthase gene (50). The degradative activity of a "bad batch" of Smal was used to make three different reading frames at lacZ and GUS. Translational and transcriptional fusions can easily be made in this vector.

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