BIO372H5 Lecture Notes - Lecture 7: Enterobacteria Phage T4, Dna Replication, Helicase
Document Summary
Many enzymes/ protein are atp dependent - capability is widespread. 30 degrees is nice temp for e coli to grow. Type 2 has some growth at 40 but plataeus after. Type 1 is deficient in helicase (at 40 degrees, e. coli didn"t grow right away) Dnab - impt in helicase, has an atpase activity. Circular dna single stranded - some of the segments will have nicks. We take probe dna and label 5" end (what kind of label, substrate - need to know this) Because of the nicks, not because of super coiling. You can measure product to have an idea how helicase is working. You have to have protein to get products. Amount of products varies dnab and ssb has an effect. Fewer copies per cell will have more activity. Stimulation of dna synthesis by t4 phage ssb, gp32. Look at short region of synthesis, and the rate of incorporation of labeled dna.
