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	<h1>'''DNA Gel Electrophoretic'''</h1>		<h1>'''DNA Gel Electrophoretic'''</h1>
−	After we link SUC2 and VP successfully, we run the PCR with an intention to confirm the expression of VP. As the figure shows, we get the genetic stripe about xxx bp which means the PCR is successful.	+	After we link SUC2 and VP successfully, we run the PCR with an intention to confirm the expression of VP. As the figure shows, we get the genetic stripe about 1101 bp which means the PCR is successful.
	[[File:Kozak_sequence.png ‎|400px|thumb|center|Figure1. This figure shows the most possible kozak consensus sequence.]]		[[File:Kozak_sequence.png ‎|400px|thumb|center|Figure1. This figure shows the most possible kozak consensus sequence.]]

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Revision as of 07:22, 20 October 2019

AOX1-Kozak-SUC2-VP-His tag-AOX1 Terminator

The SUC2 gene of S. cerevisiae, encoding invertase was designed to codon preference of P. pastoris excluding the sequence of the N-terminal pre-pro-sequence signal peptide and His-tag in the C terminus. VP can catalyze lignin.

Characterization

This is a four section for degrade and transfer lignin part.The expression of SUC2 gene is a kind of Sucrose transferase [1], and its signal peptide can help the protein out cells. We use this signal peptide raise the efficiency of VP’s transformation.

DNA Gel Electrophoretic

After we link SUC2 and VP successfully, we run the PCR with an intention to confirm the expression of VP. As the figure shows, we get the genetic stripe about 1101 bp which means the PCR is successful.

SDS-PAGE

We run the SDS-PAGE to check whether SUC2 help the enzyme transfer to the extracellular. As the figure shows, we get the protein type about xxx bp which means the SDS-PAGE is successful.

Enzyme Activity

We use manganese and hydrogen peroxide to detect the enzyme activity. As the figure shows, the solution turns xxx, which confirm the enzyme activity.

Reference

[1]Cloning and expression of Saccharomyces cerevisiae SUC2 gene in yeast platform and characterization of recombinant enzyme biochemical properties. Nooshin Mohandesi, Seyed Omid Ranaei Siadat, Kamahldin Haghbeen, Ardeshir Hesampour Received: 8 February 2016 / Accepted: 25 May 2016 / Published online: 8 June 2016 The Author(s) 2016.