Difference between revisions of "Part:BBa K3717014"
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Endo-β-galactosidase an enzyme that catalyzes the cleavage of the A and B type blood antigens (trisaccharides) such that the remaining sugar can be classified as a H antigen, which the anti-A and anti-B antibodies are unable to recognize and thus does not elicit an immune response in the human body. Thus, the enzyme can convert both A and B blood types to universal O type. The enzyme has been previously shown to successfully convert A,B, and AB type erythrocytes to O type but lacked a cost-effective method of production. | Endo-β-galactosidase an enzyme that catalyzes the cleavage of the A and B type blood antigens (trisaccharides) such that the remaining sugar can be classified as a H antigen, which the anti-A and anti-B antibodies are unable to recognize and thus does not elicit an immune response in the human body. Thus, the enzyme can convert both A and B blood types to universal O type. The enzyme has been previously shown to successfully convert A,B, and AB type erythrocytes to O type but lacked a cost-effective method of production. | ||
| − | We obtained the amino acid sequence of the endo-β-galactosidase protein, derived from Clostridium perfringens, from the iGEM DNA Repository Plate (BBa_K1483001), which served as our Open Reading Frame (ORF). We attached a T7 promoter, derived from the T7 phage, and strong ribosome binding site (RBS; BBa_K525998) upstream of the open reading frame (ORF). The composite gene was synthesized through DNA cloning. | + | We obtained the amino acid sequence of the endo-β-galactosidase protein, derived from <i>Clostridium perfringens</i>, from the iGEM DNA Repository Plate (BBa_K1483001), which served as our Open Reading Frame (ORF). We attached a T7 promoter, derived from the T7 phage, and strong ribosome binding site (RBS; BBa_K525998) upstream of the open reading frame (ORF). The composite gene was synthesized through DNA cloning. |
Revision as of 09:47, 19 October 2021
Endo-β-Galactosidase with T7 promoter and strong RBS
Figure 1: Endo-β-galactosidase from 2014 Tuebingen with T7 promoter, RBS, and a terminator.
Contruct Design
Endo-β-galactosidase an enzyme that catalyzes the cleavage of the A and B type blood antigens (trisaccharides) such that the remaining sugar can be classified as a H antigen, which the anti-A and anti-B antibodies are unable to recognize and thus does not elicit an immune response in the human body. Thus, the enzyme can convert both A and B blood types to universal O type. The enzyme has been previously shown to successfully convert A,B, and AB type erythrocytes to O type but lacked a cost-effective method of production.
We obtained the amino acid sequence of the endo-β-galactosidase protein, derived from Clostridium perfringens, from the iGEM DNA Repository Plate (BBa_K1483001), which served as our Open Reading Frame (ORF). We attached a T7 promoter, derived from the T7 phage, and strong ribosome binding site (RBS; BBa_K525998) upstream of the open reading frame (ORF). The composite gene was synthesized through DNA cloning.
References
Rahfeld, Peter, and Stephen G. Withers. “Toward Universal Donor Blood: Enzymatic Conversion of A and B to O Type.” Journal of Biological Chemistry, vol. 295, no. 2, Jan. 2020, pp. 325–34. DOI.org (Crossref), https://doi.org/10.1074/jbc.REV119.008164.
Kwan, David H., et al. “Toward Efficient Enzymes for the Generation of Universal Blood through Structure-Guided Directed Evolution.” Journal of the American Chemical Society, vol. 137, no. 17, American Chemical Society, May 2015, pp. 5695–705. ACS Publications, https://doi.org/10.1021/ja5116088.
UniProtKB - Q6RUF5 (EABC_CLOPF). UniProt, 2 June 2021, www.uniprot.org/uniprot/
Q6RUF5. Accessed 19 Oct. 2021.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]