Showing papers by "Torrey Pines Institute for Molecular Studies published in 1988"

λ ZAP: a bacteriophage λ expression vector with in vivo excision properties

Abstract: A lambda insertion type cDNA cloning vector, Lambda ZAP, has been constructed. In E. coli a phagemid, pBluescript SK(-), contained within the vector, can be excised by f1 or M13 helper phage. The excision process eliminates the need to subclone DNA inserts from the lambda phage into a plasmid by restriction digestion and ligation. This is possible because Lambda ZAP incorporates the signals for both initiation and termination of DNA synthesis from the f1 bacteriophage origin of replication (1). Six of 21 restriction sites in the excised pBluescript SK polylinker, contained within the NH2-portion of the lacZ gene, are unique in lambda ZAP. Coding sequences inserted into these restriction sites, in the appropriate reading frame, can be expressed from the lacZ promoter as fusion proteins. The features of this vector significantly increase the rate at which clones can be isolated and analyzed. The lambda ZAP vector was tested by the preparation of a chicken liver cDNA library and the isolation of actin clones by screening with oligonucleotide probes. Putative actin clones were excised from the lambda vector and identified by DNA sequencing. The ability of lambda ZAP to serve as a vector for the construction of cDNA expression libraries was determined by detecting fusion proteins from clones containing glucocerbrosidase cDNA's using rabbit IgG anti-glucocerbrosidase antibodies.

Assay and apparatus using a lateral flow, non-bibulous membrane

Abstract: A method and apparatus for conducting specific binding pair assays, such as immunoassays, is described A porous membrane capable of non-bibulous lateral flow is used as assay substrate; a member of the binding pair is affixed in an indicator zone defined in the substrate The sample is applied at a position distant from the indi­cator zone and permitted to flow laterally through the zone; any analyte in the sample is complexed by the af­fixed specific binding member, and detected A novel method of detection employs entrapment of observable particle in the complex Blood is a particularly pre­ferred sample as the red blood cells can be used as the observable particles for detection of the complex

Class I alloantigen is sufficient for cytolytic T lymphocyte binding and transmembrane signaling.

Abstract: Based largely on antibody blocking studies, a number of surface "accessory" molecules on effector cytolytic T lymphocytes (CTL) have been implicated as having a role in mediating CTL binding and lysis of target cells, possibly via binding to ligands on the target cell surface. Despite this, cloned allogeneic CTL were able to specifically bind cell-size, artificial membranes (pseudocytes) bearing only class I alloantigen. This binding triggered CTL degranulation, as measured by serine esterase release. Thus, class I alloantigen alone is both a necessary and sufficient ligand for specific binding and effective transmembrane signaling to occur.

Perturbation of the Autoimmune Network. I. Immunization with Anti-Idiotypic Antibodies Prior to Challenge with Antigen Induces Quantitative Variations in the Autoantibody Response

Abstract: The effect of immunization with anti-idiotypes on the production of autoantibodies subsequently induced with nominal antigen, thyroglobulin, was investigated in BALB/c mice with three distinct rabbit antibodies and one syngeneic monoclonal antibody (mAb) specific for an idiotype (Id62) borne on a mouse monoclonal autoantibody to thyroglobulin. In these in vivo experiments, dose, route and form of the anti-idiotypic immunization were kept constant, but the time interval between exposure to anti-idiotype and antigen challenge was varied. When the interval was short (two weeks), heterologous anti-Id62 antibodies predisposed to suppression of the autoantibody response in 2/3 instances. Suppression was also obtained in mice immunized with a syngeneic mAb directed against Id62. In each case, a prominent idiotype-positive (Id’) response was measured in the serum. Adsorption/elution studies indicated that Id’ molecules in suppressed mice were principally non antigen-binding. on the other hand, when the time inter...

Telecommunication network analysis with COMNET 11.5

Abstract: COMNET Il.5 is a general SIMSCRIPT Il.5-based model for the performance analysis of circuit-switched, message-switched, or packet-switched networks. A variety of alternate-routing and adaptive shortest-path routing algorithms are built-in. After creating a network description with a convenient menu-driven editor, simulation follows immediately. There are no programming delays. Network operation is animated during the simulation. Reports show blocking probabilities, call queueing and packet delays, network throughput, link group utilization, and link group queue statistics. This paper highlights some of COMNET Il.5's latest features, which include capabilities for modeling call preemption, call retries, dynamic bandwidth allocation, link failures, and source-node routing.