PYNR071C is the promoter of aldose 1-epimerase superfamily protein YNR071C
A promoter originates from the yeast, which could be induced by xylose to activate genes related to material transportation. When the carbon source is mainly xylose, cell prefer to absorb extracellular nutrient. Xylose is the second most abundant saccharide in cellulose-like bio-staple, such as straw, right after glucose. Engineered yeast which is able to ferment xylose effectively, could be used to increase the availability of material and decrease the cost of ethanol extraction from cellulose fuel. As a kind of safe carbon source of relatively low cost, xylose could have been used in food industries, while there is few study about its promoter at present. Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000COMPATIBLE WITH RFC
AOX1 promoter is the strongest eukaryotic promoter currently known in yeast expression system. So we choose AOX1 as the primary promoter when we synthesized all these plasmid for the sake of more convenient expression. But noticing that methanol is hazardous, flammable, combustible and therefore, inappropriate to have direct contact with the hair, we need to substrate AOX1 for constitutive promoter Panb1 and xylose induced promoter Pynr071C to realize the projected regulation function as designed. Double-enzyme cleavage and rejointing is used to achieve this. We amplify the target gene located in the primarily synthesized plasmid without AOX1 promoter, and digest the acquired fragments and two kinds of plasmid, containing promoter only, with EcoR I and BamH I, then transform them into E.coli after linking the product together. Through this, we successfully substrate the primary AOX1 for Panb1 and Pynr071C.
All the bands are identical to the theoretical lengths, which could demonstrate that these plasmid are correctly constructed and successfully transformed into E.coli, confirmed by sequencing.
To verify whether our Pynr071c promoter could initiate the expression of downstream genes, we linked GFP it, then transfect it into yeast and test the exist of fluorescence.