The specificity loop of T7 RNA polymerase interacts first with the promoter and then with the elongating transcript, suggesting a mechanism for promoter clearance
AuthorsTemiakov D, Mentesana PE, Ma K, Mustaev A, Borukhov S, McAllister WT
JournalProc Natl Acad Sci U S A
PubMed ID11095736
During the early stages of transcription, T7 RNA polymerase forms an unstable initiation complex that synthesizes and releases transcripts 2-8 nt in length before disengaging from the promoter and isomerizing to a stable elongation complex. In this study, we used RNA small middle dotprotein and RNA small middle dotDNA crosslinking ... More
Genome-scale cloning and expression of individual open reading frames using topoisomerase I-mediated ligation.
The in vitro cloning of DNA molecules traditionally uses PCR amplification or site-specific restriction endonucleases to generate linear DNA inserts with defined termini and requires DNA ligase to covalently join those inserts to vectors with the corresponding ends. We have used the properties of Vaccinia DNA topoisomerase I to develop ... More
beta -Glucoside Kinase (BglK) from Klebsiella pneumoniae. PURIFICATION, PROPERTIES, AND PREPARATIVE SYNTHESIS OF 6-PHOSPHO-beta -D-GLUCOSIDES.
ATP-dependent beta-glucoside kinase (BglK) has been purified from cellobiose-grown cells of Klebsiella pneumoniae. In solution, the enzyme (EC ) exists as a homotetramer composed of non-covalently linked subunits of M(r) approximately 33,000. Determination of the first 28 residues from the N terminus of the protein allowed the identification and cloning ... More