High expression of human chorionic gonadotrophin beta-subunit using a synthetic vaccinia virus promoter.
We have constructed a recombinant vaccinia virus to express the beta-subunit of human chorionic gonadotrophin (betahCG), a secretory glycoprotein that is used as an antigen for a contraceptive vaccine. The cDNA encoding the subunit was cloned under the control of a synthetic promoter that could be recognised by a vaccinia virus RNA polymerase to direct transcription. The peak expression level of betahCG directed by a late synthetic promoter (Psyn) was 11.5 microg/ml, a level that was at least sixfold higher than that directed by the p7.5 early/late promoter. The expressed protein was correctly processed post-translationally such that it attained a conformation with correctly folded discontinuous epitope(s) similar to that seen in native betahCG.
Gupta
A
A
National Institute of Immunology, New Delhi 110067, India.
Chandrasekhar
S
S
Pal
R
R
Ahlawat
S
S
Singh
O
O
D015139:Blotting, Southern; D002460:Cell Line; D018997:Chorionic Gonadotropin, beta Subunit, Human / Q000235:genetics*; D006801:Humans; D010446:Peptide Fragments / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D014616:Vaccinia virus / Q000235:genetics*
D018997:Chorionic Gonadotropin, beta Subunit, Human; D010446:Peptide Fragments; C063554:urinary gonadotropin fragment