Team:GDSYZX/Results

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Result

First, we got the gene wild type pcs1 from the arabidopsis cDNA. Then we use ‘BamH 1’ and ‘Not 1’as the restriction enzyme to cut the wild type pcs1 and pEASY-Blunt, a kind of cloning vector. And then we linked them and transformed them to DH5α. Here is the colony PCR with it. Then, we connected wtpcs1 with pPIC9K and transformed them into BL21. The next picture is the plasmid of the pPIC9K-wtpcs1.The band is the location of pPIC9K-wtpcs1.

We also did the same things with copcs1. The first picture is the PCR of copcs1. The next picture is the colony PCR of pEASY-copcs1 in DH5α. The last picture is the plasmid of pPIC9K-copcs1.

Furthermore, we transformed the pPIC9K-wtpcs1 and pPIC9K-copcs1 into the GS115. And then we cultured them in MD culture medium without histidine. Here is the picture.

And we extracted the protein of prokaryotic expression, including the wtpcs1 and copcs1. We used the IPTG to induce BL21 to express protein. We divided them into 4 groups. And then we used IPTG to induce them for 4 hours, 6 hours, 8 hours, 12 hours，respectively. There is the result. If we obtained the protein, it would locate in 54 kD. However, the protein can’t be express, since pcs1 is a eukaryotic gene.

In addition, we have done the experiment about yeast’s tolerance to cadmium chloride. We divided them into six groups. There were added 100 microliters from 500 micro mole per liter to 1000 micro mol per liter. There is a gap of 100 between each group. We made three copies for each group in order to make the data be more convictive. The picture shows that yeast can’t survive in 900 micro mole per liter.

Furthermore, we have done some modeling. First of all, we attempt to accomplish the yeast growth curve. Look at the graph. We diluted the yeast solution at seven different levels and make three copies for each one. And then, we calculate the mean.

Then, we calculate the regression curve. The R-square is equal to 0.9954.

Secondly, we try to find out the relation between cadmium concentration and yeast concentration. We cultured them in different concentration in 1 micro liter. We measure the od value and compute the yeast concentration from the yeast growth curve.

Here is the scatter diagram. It seems that the second point is outlier.

Deleting the outlier, we obtained the regression curve.

Eventually, we make another group to find out the relation in 5 micro liters. We change the cadmium concentration and draw a new graph. The second point is a outlier. Canceling the outlier, we gain the regression curve.