Mechanism II

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Yeast can produce riboflavin itself, but it can only supply its own metabolism without accumulation. There are 6 genes related to the riboflavin production. The patyway of riboflavin synthesis is showed below:

Figure 4 the riboflavin synthesis pathway

We propose to change the prmoters related to riboflavin synthesis of Yeast to strong promter ADH1. The two-step-gene replacement method is used in our project. Take RIB1 as an example. Take the RIB1gene as three regions: region1, region2 and region2 (figure5)

Figure 5 the regions in RIB1 gene
Figure 6 plasmid Yiplac211

Region 2 contains the RIB1 original promoter. Region3 is a part of gene ecoding region and region 1 is the upstream region of RIBI original promoter. Our aim is to substitude ADH1 for region2. We take the ADH1 strong promoter as region4. The plasmid used in this experiment is YIPlac211(figure 6). The experimental process is:

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