Difference between revisions of "Part:BBa K3717014"

Revision as of 08:39, 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.

<b>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.

Sequence and Features