Enhancement of Defective RNA Expression Vectors as Potential Vaccine Delivery Systems for Avian Infectious Bronchitis Virus
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IBV defective RNA (D-RNA) CD-61, a deletion mutant of the naturally occurring D-RNA CD-91, is capable of being utilised as an RNA expression vector for heterologous genes (Pénzes et al., 1996; Stirrups et al., 2000a; Stirrups et al., 2000b). The reporter genes chloramphenicol acetyltransferase (CAT) and luciferase, under the control of IBV gene 5 transcription associated sequence (TAS), are capable of expression from within CD-61 during D-RNA rescue. However the modification of CD-61 by the insertion of foreign gene sequences has an adverse effect on D-RNA rescue efficiency resulting in an eventual loss of foreign gene expression (Stirrups et al., 2000b). We attempted to enhance heterologous gene expression in CD-61 by utilising the encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES) as an alternative to gene 5 TAS to control CAT gene expression.
KeywordsInternal Ribosome Entry Site Cell Free System Infectious Bronchitis Virus Rabbit Reticulocyte Lysate Heterologous Gene Expression
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