Lab Board

Plant transformation of agro gul4 cDNA construct.

Puna Maya Maharjanl 2011-02-07l Hit 646

Last week,

1. I did completed the western blot of T2 of 35S:CYP83B1:YFP:HA in ws-2 and gul4 background.

2.

a. To confirm the transformation is ok,I did PCR of T1 of Promoter CYP83B1gDNA:GUS in Ws-2 and gul4 background (12 lines from each) with one primer form gene and anothe from HA tag.

   b. I did take photo of those lines.

c.  Since the clear phenotype complementation is not seen in gul4 background I wanted to select more lines. So, I sowed more To seeds in hygromycene media.

3. Plant transformation of agro of 35S: gul4 cDNA:YFP:HA in gul4 and Ws-2 backgrounds for 1st and second time.

4. Photography of 8 weeks old Ws-2, gul4, Col, atr4-2 and atr4-1.

5.GUS staining of 10 lnes of T2 of at4g31250 promoter gDNA:GUS in Ws-2 backgrounds. 3 liones of them showed strong staining in elongation and dividng zone of root. 2 lines showed moderate staining and the rest of the lines showed weak staining. So I transplanted about 8 to 10 seedlings from 3 strong lines and 1 moderate lines into soils.

6. I sowed the seeds of F1 of atr41Xgul4. gul4Xatr4-2. gul4Xatr4-2and atr4-2Xgul4. to see whether they segregate or not.

7.Ws-2, col, gul4 and atr-2 seeds sown for seedling phenotype in light and dark.

 Today,

I will do RNA isolation of dose response treatment of BL in Ws-2 and if time permits continue to cDNA synthesis.