Chloramphenicol acetyltransferase reporter gene

Figure 3 Schematic diagram of an expression plasmid containing the kanamycin resistance gene, origin of bacterial replication, cytomegalovirus immediate-early promoter enhancer (CMV), ubiquitin B promoter and intron, chloramphenicol acetyltransferase reporter gene (CAT), and bovine growth hormone (BGH) polyadenylation signal.

This chapter covers j8-galactosidase (jSGal or LacZ) and luciferase (Lux/Luc) gene systems as typical reporter systems. Note that alkaline phosphatase (PhoA) can also be used as an excellent reporter system and has been described in Chapter 5. As selection markers, the ampicillin resistance system (Bla or ApO will be described. The chloramphenicol acetyltransferase (CAT) gene system will also be discussed as a typical marker/reporter with dual functions. 

They used different generations of intact dendrimers to transfect plasmid DNA in a variety of cells (Table 18.2) using luciferase, CAT (chloramphenicol acetyltransferase) and /i-galactosidasc reporter genes to quantify transfection efficiency. 

There are other reporter gene systems, such as j8-galactosidase (a bacterial enzyme), chloramphenicol acetyltransferase (a bacterial enzyme), and aequorin (a jellyfish protein). 

DNA injection directly into mouse diaphragm has also resulted in luciferase expression and there appeared to be no damage to the diaphragm due to the DNA injections (Davis and Jasmin, 1993). In a related study, /3-galactosidasc ( /3-gal)-encoding pDNA injected into the articular space of rabbit knee joints resulted in /3-gal expression in the joints (Yovandich etal., 1995). In the same study, chloramphenicol acetyltransferase (CAT) encoding pDNA injected into rat knee joints also led to reporter gene expression, with peak expression 48 hours after injection and with no detectable activity 15 days later. 

---