title: Replacement of the human cytomegalovirus promoter with fish enhancer and core elements to control the expression of the G gene of viral haemorrhagic septicemia virus (VHSV).
abstract: This work explores some of the possibilities to replace human cytomegalovirus (CMV) core and/or enhancer promoter control elements to create new expression vectors for use with fish. The work is relevant to fish vaccination, since DNA vaccines use eukaryotic expression plasmids controlled by the human cytomegalovirus (CMV) promoter to be effective against novirhabdoviruses, such as viral haemorrhagic septicemia virus (VHSV), one of the most devastating fish viral European diseases. To reduce possible homologous recombination with fish genome, core and enhancer sequences from fish origin, such as trout interferon-inducible myxovirus protein (Mx), zebrafish retrovirus long terminal repeat (LTR) and carp β-actin (AE6), were combined with those of CMV to design alternative hybrid promoters. The substitution of CMV core and/or enhancer with the corresponding elements of Mx or the LTR core maintained a similar in vitro protein G expression level than that obtained by using the CMV promoter. Vectors using the dsRNA-inducible Mx enhancer followed either by the LTR or the AE6 cores showed the highest in vitro protein G expression levels. Furthermore, synthetic constructs using the Mx enhancer maintained their polyI:C induction capabilities despite the core used. Some of these hybrid promoters might contribute to the development of all-fish-vectors for DNA vaccines while others might be useful for more basic studies.
authors.affiliation: IBMC Universidad Miguel Hernández, Alicante, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Martinez-Lopez
authors.forename: B
authors.initials: B
authors.lastname: Chinchilla
authors.forename: P
authors.initials: P
authors.lastname: Encinas
authors.forename: E
authors.initials: E
authors.lastname: Gomez-Casado
authors.forename: A
authors.initials: A
authors.lastname: Estepa
authors.forename: J M
authors.initials: JM
authors.lastname: Coll
mesh_terms: D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D003587:Cytomegalovirus / Q000235:genetics*; D029941:Fish Proteins / Q000235:genetics*; D005399:Fishes; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D029941:Fish Proteins; C096089:G protein, viral hemorrhagic septicemia virus; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins