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Genetic element
(GENE)
last updated: 21 Jul 2026
Synthetic subtilisin
EN
-
TUBSUB1EN
CS-TUBSUB1-SYNTH
EN
Protein coding sequence
Yes
The TUBSUB1 coding sequence from Toxoplasma gondii was redesigned using codons preferentially used by Bacillus subtilis to improve protein expression while preserving the amino acid sequence of the antigen. The optimized gene was subsequently synthesized chemically prior to cloning into the expression plasmid.
Subtilisin
EN
Rather than selecting the Bacillus subtilisin protein (serine endopeptidase), a conserved protein that could serve as a pan-Apicomplexa antigen from a previously characterized in Toxoplasma gondii was used an parasite closely related to Eimeria. The protein was predicted to be associated with the parasite microneme, an organelle involved in host-cell invasion of coccidia parasites which Eimeria is one of the primary causative agents. Subtilisins are widely used as engineered enzymes for industrial applications and as secretion tags to efficiently produce and export foreign proteins out of bacterial cells. Subtilisin-like proteases in Eimeria are believed to play essential roles during the parasite life cycle by participating in host cell invasion and by proteolytically processing precursor proteins (e.g., GAM56 and GAM82) required for formation of the resilient oocyst wall. This processing enables the incorporation and cross-linking of tyrosine-rich peptides into the oocyst wall, and disruption of these steps is predicted to impair oocyst development and parasite survival.
EN
- Subtilisin is a native extracellular serine protease produced by Bacillus subtilis that degrades extracellular proteins. For this reason, many industrial B. subtilis production strains carry deletions or mutations of the genes that encodes subtilisin to reduce extracellular protease activity and degradation of recombinant vaccine antigens and to improve recombinant protein yield and stability. While subtilisin is primarily known as a powerful serine protease secreted by B. subtilis and subtilisin-like proteases are critical for these essential biological processes, subtilisin-like proteases from pathogens are often investigated as vaccine antigens because they are exposed on the pathogen surface or are involved in host invasion, cutting peptide bonds into usable vaccine antigens.
- BIOTECH VAC COX uses a modified Bacillus subtilis strain to express a recombinant Eimeria antigen that elicits an immune response against the parasite following oral administration to birds.
- The recombinant protein is described as a synthetic, codon-optimized gene encoding a conserved Eimeria antigen, derived by homology from a Toxoplasma gondii protein. The coding sequence of Toxoplasma gondii subtilisin was used to identify its orthologous protein in Eimeria spp. The gene sequence was then designed in silico (GenScript) for Bacillus subtilis codon-optimization in order to encode an Eimeria antigen.
- The synthetic subtilisin TUBSUB1 was inserted to plasmid pHT01 and transformed into Escherichia coli DH5α (DH5 alpha). The plasmid from the resulting E. coli transformant was purified and used for transformation into the B. subtilis donor organism.
- The TUBSUB1 listed as the protein coding sequence name is provided by the submitter. Should there be another name to the coding sequence or any additional information regarding the genetic element, the record will be updated.
| Record type | Field | Record(s) | |
|---|---|---|---|
| Living Modified Organism | Introduced or modified genetic element(s) | 1 | |
