Team:HUST-China/Proof Of Concept

Proof Of Concept | iGEM HUST-China

Proof Of Concept


Introduction

Our project (click design for detailed information) integrated at most five reactions within a system. Each reaction in the system is interrelated by metabolic or regulatory pathways. Integrated with the systems, the Pichia pastoris GS115 can express target products which are separated and applied with our hardware. We will prove the validity and feasibility from the following three aspects including construction, expression and application.

Construction

Every system is composed of dye and perm device as well as decolorization and straightening device, constituted of several parts. The construction process shall be divided into two parts: first connecting the parts to build devices, next assembling devices into systems. During this process, we chose seamless cloning of homologous recombination at the beginning and turned to enzyme digestion and ligation, finally getting the expected result through the latter method.

Expression

The constructed devices and systems were transformed into pichia pastoris GS115 and expressed. We added α-factor as the signal peptide in front of the functional genes for extra-cellular secretion but ended up unsatisfactory. The signal peptide was removed for intracellular expression and we got ideal result, with most protein successfully expressed. The activity of two kinds of enzymes were measured.

Application

When it comes to the application of our products, we should ensure the theoretical basis of our pathways. Furthermore, to put it into the real world for use, we need the support of hardware. In this section, we introduce these two aspects.

Hmm…I can hardly pick out any flaw in this part, what about you? Here is another thing I really care about—the safety of the products! Don’t you worry about whether the products of this team can cause cancer or do damage to hair like other hairdressing products in the market?

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