1. Assume that a 4000-bp long circular DNA plasmid has two HindIII sites at coordinate 1 and 801.  If we digest this circular DNA plasmid with BamHI, we obtain 500 bp, 1400 bp, and 2100 bp DNA fragments.  If we perform HindIII/BamHI double digestion with this circular plasmid, we obtain 200 bp, 500 bp, 600 bp, 1200 bp, and 1500 bp DNA fragments.  From above information, please interpret the location of BamHI sites.

(1)  How many BamHI sites are there in this circular plasmid?

(2)  Where are those BamHI sites? Please draw the map.

2. Design the primers to clone the specific gene into the vector as indicated to make the fusion protein with FLAG tag.  You will need to show the Tm, primers, restriction enzymes, and recombinant DNA molecule.  Please follow the instruction to find your project.

            pESCTRP       ESA1               (your last digit of SS# is 0 &1)

            pESCHIS        INO80             (your last digit of SS# is 2 &3)

            pESCLEU       HHT1              (your last digit of SS# is 4 &5)

            pESCHIS        HHO1              (your last digit of SS# is 6 &7)

            pESCLEU       HHF1              (your last digit of SS# is 8 &9)

3. Please describe one example (including the experiments) to demonstrate that the H1 may be repressor of specific genes.

4. Please describe one example (including the experiments) to demonstrate that the histone acetylation is important to transcriptional activation.