Influence of codon usage and translational initiation codon context in the AcNPV-based expression system: computer analysis using homologous and heterologous genes.
Codon usage by all the known gene sequences from Autographa californica nuclear polyhedrosis virus (AcNPV) was compared with that of firefly luciferase (luc) and the beta subunit of human chorionic gonadotropin (beta hCG) expressed to contrasting levels in the baculovirus system. The highly expressed luc gene showed a codon usage similar to AcNPV genes, as reflected by a very low D-squared statistic value (0.78) and a similar G/C usage (45%) at wobble positions. However, the underexpressed beta hCG gene displayed a high D-squared value (7.3) and G/C usage (82.5%) at the wobble base position. Alignment of the 20 nucleotides around the initiation codon of 23 AcNPV genes identified a novel consensus translation initiation sequence aag/ta/tat/aa/cAAaATGaa/ct/ag/aAan, which was quite different from the Kozak consensus sequence (GCC)GCCA/GCCATGG. An extension of these analyses to a sample of other heterologous genes overexpressed and underexpressed in BEVS suggested similar trends. These theoretical analyses have important implications for heterologous gene expression in this system.
Ranjan
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Eukaryotic Gene Expression Laboratory, National Institute of Immunology, New Delhi-India.
Hasnain
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D000818:Animals; D001483:Base Sequence; D006063:Chorionic Gonadotropin / Q000235:genetics; D018997:Chorionic Gonadotropin, beta Subunit, Human; D018387:Codon, Initiator; D004279:DNA, Viral; D001330:Electronic Data Processing; D015967:Gene Expression Regulation, Viral; D006801:Humans; D008156:Luciferases / Q000235:genetics; D008969:Molecular Sequence Data; D009036:Moths; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000378:metabolism; D010446:Peptide Fragments / Q000235:genetics; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000235:genetics
D006063:Chorionic Gonadotropin; D018997:Chorionic Gonadotropin, beta Subunit, Human; D018387:Codon, Initiator; D004279:DNA, Viral; D010446:Peptide Fragments; D011994:Recombinant Proteins; D008156:Luciferases