Polynucleotides. CXXIII. Physical characterization and simultaneous purification of bacteriophage T4 induced polynucleotide kinase, polynucleotide ligase, and deoxyribonucleic acid polymerase

Nucleotide sequence around the replication origin of polyoma virus DNA

“…and then these DNA fragments were labelled with a2p at the 5'.termini using T4-induced polynucleotide kinase [11 ] and [~/-a2P]ATP [12]. The labelled fragments were digested by HhaI endonuclease and the digests separated by electrophoresis in an 8% polyacrylamide gel.…”

DNA ligases of eukaryotes

“…A crude cell extract of TCinfected E. coli has about 500 times higher DNA ligase activity than the most active mammalian cell extracts under optimal assay conditions [25], and homogeneous T4 DNA ligase is an approx. 2000-fold purified enzyme [37,38]. However, it should not be concluded from such data that it might be necessary to purify the DNA ligase I about lo6 times in order to obtain a homogenous enzyme.…”

Enzymatic cleavage of m7 GDP from eucaryotic mRNA

“…Like mRNAs of prokaryotes and poliovirus, decapped mRNAs produced by polynucleotide kinase have a single phosphate group at the 5'-terminus. The enzyme can be purified to homogeneity free of nucleases [8] . Therefore this enzyme can be a useful tool for comparing the biological activity of capped and decapped mRNA.…”

“…Bacteriophage Tq-induced polynucleotide kinase isolated according to Panet et al [8] was used to cleave the cap structure. The reaction mixture contained per 0.2 ml: 65 mM Tris-HCl (pH 7.8) 15 mM mercaptoethanol, 9 mM magnesium chloride, 30 pg mRNA or cap structures isolated from 50 pg mRNA, and 3 units of highly purified polynucleotide kinase.…”