Torrey Pines Institute for Molecular Studies

About: Torrey Pines Institute for Molecular Studies is a nonprofit organization based out in San Diego, California, United States. It is known for research contribution in the topics: Antigen & T cell. The organization has 2323 authors who have published 2217 publications receiving 112618 citations.

Polishing with T4 or Pfu polymerase increases the efficiency of cloning of PCR fragments

Abstract: Several of the polymerases widely used for PCR, such as Taq DNA polymerase, have been found to exhibit terminal deoxynucleotide transferase activity (1, 2, 3). This terminal transferase activity, for the most part is limited to the addition of a single nucleotide and has been named 'extendase' activity. In a widely cited study, Clark has shown that when the 3' base is a cytosine nucleotide, the Taq DNA polymerase adds predominately a single A base. This result forms the basis for the T/A cloning procedure. Although Clark's results were true for 3' C-ending fragments, Hu (3) has shown that the extended nucleotide is dependent on the specific nucleotide present at the 3' end of the synthetic double-stranded polydeoxyoligonucleotide. Different polymerases, such as Taq, T7, Klenow and Vent have different extendase characteristics with regards to which base is added at the 3' DNA ends. The polymerases derived from T4 bacteriophage and the hyperthermophilic archea Pyrococcus furiosus (available from Stratagene, La Jolla, CA) do not contain any extendase activity and in all cases were found to leave bluntended molecules.

Parallel Array and Mixture-Based Synthetic Combinatorial Chemistry: Tools for the Next Millennium

Abstract: Technological advances continue to be a central driving force in the acceleration of the drug discovery process. Combinatorial chemistry methods, developed over the past 15 years, represent a paradigm shift in drug discovery. Initially viewed as a curiosity by the pharmaceutical industry, combinatorial chemistry is now recognized as an essential tool that decreases the time of discovery and increases the throughput of chemical screening by as much as 1000-fold. The use of parallel array synthesis approaches and mixture-based combinatorial libraries for drug discovery is reviewed.

Palmitoylation of the Cytoplasmic Domain of the Neural Cell Adhesion Molecule N-CAM Serves as an Anchor to Cellular Membranes

Abstract: The neural cell adhesion molecule N-CAM is expressed at key sites during embryonic development and mediates homophilic adhesion between cells both in the embryo and in the adult. N-CAM is expressed in multiple forms and two of the major isoforms differ in their cytoplasmic domains, one (Id form) having an insert of 261 amino acids that is missing in the other (sd form). N-CAM has been previously shown to be palmitoylated. but the sites of acylation have not been localized. We show here that the cytoplasmic domain of the N-CAM became palmitoylated after transfection of a cDNA encoding N-CAM into COS-7 cells, and that this acylation occurs on the four closely spaced cysteines in the cytoplasmic domain of N-CAM. Moreover, when a cDNA encoding only the cytoplasmic domain was transfected into cells, the protein was palmitoylated and associated with membranes even though it lacked a membrane spanning segment. Site directed mutagenesis of the four cysteine residues to serines at positions 5. 11. 16, and 22 in th...