Team:Saint Joseph/Parts

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Parts

We added four basic parts to the IGEM registry: BBa_K3941000, BBa_K3941001, BBa_K3941002, BBa_K3941003.

BBa_K3941000 is a codon-optimized CelAB's catalytic region's sequence for E. coli DH5⍺ with histidine tag at the end for the ease of protein isolation. The CelAB which is produced by T. turnerae T7902, an endosymbiont of the shipworm Lyrodus pedicellatus. CelAB were examined to evaluate its potential multiple enzymatic activities and carbohydrate binding affinities. We conducted a spectrophotometer absorbance analysis but the numbers that we obtained were too high.

The second basic part that we designed is BBa_K3941001 which originated from endoglucanase II (EGII) gene which breaks down beta-1,4-glycosidic bonds in cellulose.

The third basic part, BBa_K3941002, is an endoglucanase II (EGII) gene which contains a mutation to form a different amino acid. We changed the amino acid Cystine from 99th position to Valine.

The fourth basic part, BBa_K3941002, is also an endoglucanase II (EGII) gene which contains a mutation. Whose mutation is changing Aspartic acid from the 185th position to Serine but we weren’t successful with this part.

Number Name Design
BBa_K3941000 CelAB CelAB (Codon Optimized), His-tag
BBa_K3941001 EGII CelAB (Codon Optimized), His-tag
BBa_K3941002 EGII(C99V) CelAB (Codon Optimized), His-tag
BBa_K3941003 EGII(D185S) CelAB (Codon Optimized), His-tag

Composite Parts

Number Name Design
BBa_K3941004 pET29b+CelAB CelAB (Codon Optimized), His-tag
BBa_K3941005 pET29b+EGII CelAB (Codon Optimized), His-tag
BBa_K3941006 pET29b+EGII(C99V) CelAB (Codon Optimized), His-tag
BBa_K3941007 pET29b+EGII(D185S) CelAB (Codon Optimized), His-tag