Thorkild Christensen

Learn More
The areA gene of Aspergillus oryzae was cloned by cross-hybridization with the Aspergillus nidulans areA gene and was found to encode an 866-amino-acid protein that is very similar to other fungal nitrogen regulatory proteins. The A. oryzae areA gene can complement A. nidulans areA loss-of-function mutations. Functional analyses indicated that the(More)
Domain VI at the 3' end of the 23 S ribosomal RNA from Escherichia coli was prepared using the in vitro T7 RNA polymerase system. Its structure was examined by probing with ribonucleases and chemical reagents, including a psoralen derivative, of various nucleotide specificities, using a reverse transcriptase procedure for analysis. The data provided support(More)
(15)N NMR relaxation parameters and amide (1)H/(2)H-exchange rates have been used to characterize the structural flexibility of human growth hormone (rhGH) at neutral and acidic pH. Our results show that the rigidity of the molecule is strongly affected by the solution conditions. At pH 7.0 the backbone dynamics parameters of rhGH are uniform along the(More)
Ribosomal proteins L10 and L12 are encoded in the L10 operon, situated at position 89.5 min on the Escherichia coli genetic map, and are able to regulate their own translation. The two proteins form a L10-L12 complex that is able to bind specifically to the leader sequence of the L10 operon mRNA and prevent translation. We show that the leader sequence: (i)(More)
The ability of a new class of metal binding tags to facilitate the purification of recombinant proteins, exemplified by the tagged glutathione S-transferase and human growth hormone, from Escherichia coli fermentation broths and lysates has been further investigated. These histidine-containing tags exhibit high affinity for borderline metal ions chelated to(More)
We report a novel enzymatic method by which an E. coli produced precursor to human growth hormone (hGH) can be converted to authentic hGH. The hGH precursor contains an aminoterminal extension that is removed in vitro by the enzyme dipeptidylaminopeptidase I (DAP I). The aminoterminal extension has been tailored to allow the use of very specific(More)
The complete enzymatic removal of affinity tags from tagged recombinant proteins is often required but can be challenging when slow points for cleavage exist. This study documents a general approach to remove N-terminal tags from recombinant proteins specifically designed to be efficiently captured by IMAC resins. In particular, site-directed mutagenesis(More)
In this study, integration of three discrete process aspects of the IMAC purification of Escherichia coli expressed recombinant proteins has been investigated. To this end, novel N-terminally tagged human growth hormone variants (tagged-vhGHs) have been expressed in E. coli by tank fermentation and captured directly from the cell lysate by a new IMAC(More)
We have carried out measurements of the stable binding of the ribosomal protein (r-protein) complex L10-L7/L12 to mutant forms of the mRNA leader of the rplJ operon of Escherichia coli. One of the point mutations, base 1548, which lies within the L10-L7/L12-protected region, almost completely abolishes in vitro formation of a stable complex of L10-L7/L12(More)
  • 1