Team:Shenzhen SFLS/Improvements

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Improvements

As our project requires the protein to exhibit high strength and viscidity, we decided to improve an existing part to achieve this aim.

The existing part we are selecting to improve is BBa_K197018. This part originally controls the expression of the protein mgfp5 in E. coli. However, as addressed in the introduction, mgfp5 exhibits perfect adhesion properties but is deficient in cohesion properties and strength.

[2]

Therefore, we are looking for a protein that has adequate strength as well as bio-compatibility. Here, a protein MasP1 attracted our attention.

The protein MasP1 is a component of silk spidroin, who has great bio-compatibility. Pulakesh Aich and a few co-authors introduce the characteristic of this protein and published a method of fusing mfp1-6 with MasP1[1].

Consider the similarity between mfp1-5 and the recombinant protein mgfp5, this fusing method is well worth trying on BBa_K197018 to improve it.

Therefore, we derive the sequence of the protein from the same article introduced previously and use the linker GGGGS- as depicted in the picture to create a fused protein of mgfp5 and MasP1.

[3]

The final part is loaded onto the plasmid backbone pET28a.

Observing the plasmid, the mgfp5-masp1 part formed an open reading frame of almost 900 bp. A his-tag is placed at the end of the protein to ensure the further purification.

Source:

[1]: Self-assembled adhesive biomaterials formed by a genetically designed fusion protein

Pulakesh Aich,‡a Jaeyeon An,‡b Byeongseon Yang,‡c Young Ho Ko,a Junghyun Kim, a James Murray,a Hyung Joon Cha, *c Joon Ho Roh,*a Kyeng Min Park *a and Kimoon Kim *ab

Chem. Commun., 2018, 54, 12642

Received 14th September 2018, Accepted 18th October 2018

DOI: 10.1039/c8cc07475e

rsc.li/chemcomm

[2]: Chem. Commun., 2018, 54, 12642

Received 14th September 2018, Accepted 18th October 2018

DOI: 10.1039/c8cc07475e

rsc.li/chemcomm Fig1

[3]: Chem. Commun., 2018, 54, 12642

Received 14th September 2018, Accepted 18th October 2018

DOI: 10.1039/c8cc07475e

rsc.li/chemcomm

supplementary informationFfig 1