Difference between revisions of "Part:BBa K3717013"

Revision as of 01:57, 19 October 2021

α-N-Acetylgalactosaminidase with T7 + RBS, C-Terminal 6x His-Tag, and Double Terminator

The composite part utilizes a T7 promoter + RBS (BBa_K525998), α-N-Acetylgalactosaminidase with C-Terminal 6x Histidine tag (BBa_K3717016), and a double terminator (BBa_B0015).

Construct Design

Figure 1: T7 promoter with RBS, alpha-N-Acetylgalactosaminidase, GS linker, C-terminal histidine tag, and a double terminator

α-N-acetylgalactosaminidase is an enzyme that catalyzes the cleavage of the N-acetylgalactosamine off of A type blood antigens such that the remaining sugar can be classified as an 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 A blood types to universal O type. While the enzyme has been previously shown to successfully convert A type erythrocytes to O type, it was inefficient in doing so, and lacked a cost-effective method of production.

We obtained the amino acid sequence of the α-N-acetylgalactosaminidase protein, derived from Elizabethkingia meningoseptica, from the iGEM DNA Repository Plate, which served as our Open Reading Frame (ORF). We attached a T7 promoter, derived from the T7 phage, strong ribosome binding site (RBS;BBa_K525998) upstream of the open reading frame (ORF), and a 6x Histidine tag through a flexible Glycine-Serine linker in the open reading frame but downstream of the α-N-acetylgalactosaminidase sequence for protein purification purposes. The composite gene was synthesized through DNA sequencing.

b> Characterization </b>

Sequence and Features