Abstract: The invention provides methods employing iterative cycles of recombination and selection/screening for evolution of whole cells and organisms toward acquisition of desired properties. Examples of such properties include enhanced recombinogenicity, genome copy number, and capacity for expression and/or secretion of proteins and secondary metabolites.

Abstract: A method for DNA reassembly after random fragmentation, and its application to mutagenesis of nucleic acid sequences by in vitro or in vivo recombination is described. In particular, a method for the production of nucleic acid fragments or polynucleotides encoding mutant proteins is described. The present invention also relates to a method of repeated cycles of mutagenesis, shuffling and selection which allow for the directed molecular evolution in vitro or in vivo of proteins.

Abstract: A virion expression system for a desired protein packaged in an envelope derived from a retrovirus useful in administering proteins which cross cell membranes in order to serve their function. Preferred virions are those that carry an RNA sequence that encodes cytokines or lymphokines, and includes IL-2, multiple drug resistance protein, and TNF.

Abstract: Cells producing recombinant retroviral particles are provided. The cells contain a first vector having a coding region encoding retroviral LTRs and a packaging signal under the control of an expression control system, a tRNA binding site located upstream from the packaging signal and origin of second strand DNA synthesis located downstream from the packaging signal. The cells also contain a second vector having a coding region encoding retroviral capsid proteins gag and pol under the control of an expression control system and a third vector having a coding region encoding a simian type D retrovirus envelope glycoprotein under the control of an expression control system.

Abstract: A virion expression system for a desired protein packaged in an envelope derived from a retrovirus useful in administering proteins which cross cell membranes in order to serve their function. Preferred virions are those that carry an RNA sequence that encodes cytokines or lymphokines, and includes IL-2, multiple drug resistance protein, and TNF.

Abstract: Recombinant retroviruses carrying a vector construct capable of preventing, inhibiting, stabilizing or reversing infectious, cancerous or auto-immune diseases are disclosed. More specifically, the recombinant retroviruses of the present invention are useful for (a) stimulating a specific immune response to an antigen or a pathogenic antigen; (b) inhibiting a function of a pathogenic agent, such as a virus; and (c) inhibiting the interaction of an agent with a host cell receptor. In addition, eucaryotic cells infected with, and pharmaceutical compositions containing such a recombinant retrovirus are disclosed. Various methods for producing recombinant retroviruses having unique characteristics, and methods for producing transgenic packaging animals or insects are also disclosed.

Abstract: A virion expression system for a desired protein packaged in an envelope derived from a retrovirus useful in administering proteins which cross cell membranes in order to serve their function. Preferred virions are those that carry an RNA sequence that encodes cytokines or lymphokines, and includes IL-2, multiple drug resistance protein, and TNF.

Abstract: Tumor Infiltrating Lymphocyte (TIL) cells transformed with exogenous DNA encoding tumor necrosis factor (TNF) prohormone variants are disclosed. Among such variants are the TNF .gamma. sig construct, which facilitates expression of the mature TNF polypeptide, and the noncleavable cytotoxic prohormone variants TNF.DELTA.(1.fwdarw.12) and TNF(1+12). The transformed TIL cells are useful in antitumor therapy.

Abstract: A virion expression system for a desired protein packaged in an envelope derived from a retrovirus useful in administering proteins which cross cell membranes in order to serve their function. Preferred virions are those that carry an RNA sequence that encodes cytokines or lymphokines, and includes IL-2, multiple drug resistance protein, and TNF.

Abstract: Host cells may be treated for an infection or a hyperproliferative disorder which is characterized by the presence, in the affected cells, of a trans-acting factor capable of regulating gene expression by inserting into the cells a polynucleotide construct having a cis-acting regulatory sequence which is regulated by the trans-acting factor and an effector gene which renders said cell susceptible to protection or destruction. For example, the cis-acting region may be homologous to the HIV tar region, and the effector gene may encode ricin A or HSV-1 thymidine kinase. Upon infection with HIV, the HIV tat protein activates the tar region, and induces transcription and expression of ricin A, resulting in cell death, or of HSV-1 tk, resulting in cell death upon treatment with dideoxynucleoside agents such as acyclovir and gancyclovir.

Abstract: Recombinant retroviruses carrying a vector construct capable of preventing, inhibiting, stabilizing or reversing infectious, cancerous or auto-immune diseases are disclosed. More specifically, the recombinant retroviruses of the present invention are useful for (a) stimulating a specific immune response to an antigen or a pathogenic antigen; (b) inhibiting a function of a pathogenic agent, such as a virus; and (c) inhibiting the interaction of an agent with a host cell receptor. In addition, eucaryotic cells infected with, and pharmaceutical compositions containing such a recombinant retrovirus are disclosed. Various methods for producing recombinant retroviruses having unique characteristics, and methods for producing transgenic packaging animals or insects are also disclosed.

Abstract: A virion expression system for a desired protein packaged in an envelope derived from a retrovirus useful in administering proteins which cross cell membranes in order to serve their function. Preferred virions are those that carry an RNA sequence that encodes cytokines or lymphokines, and includes IL-2, multiple drug resistance protein, and TNF.

Abstract: The present invention provides compositions and method,, for utilizing recombinant alphavirus vectors.

Abstract: Host cells may be treated for an infection or a hyperproliferative disorder which is characterized by the presence, in the affected cells, of a trans-acting factor capable of regulating gene expression by inserting into the cells a polynucleotide construct having a cis-acting regulatory sequence which is regulated by the trans-acting factor and an effector gene which renders said cell susceptible to protection or destruction. For example, the cis-acting region may be homologous to the HIV tar region, and the effector gene may encode ricin A or HSV-1 thymidine kinase. Upon infection with HIV, the HIV tat protein activates the tar region, and induces transcription and expression of ricin A, resulting in cell death, or of HSV-1 tk, resulting in cell death upon treatment with dideoxynucleoside agents such as acyclovir and gancyclovir.

Abstract: Recombinant retroviruses carrying a vector construct capable of preventing, inhibiting, stabilizing or reversing infectious, cancerous or auto-immune diseases are disclosed. More specifically, the recombinant retroviruses of the present invention are useful for (a) stimulating a specific immune response to an antigen or a pathogenic antigen; (b) inhibiting a function of a pathogenic agent, such as a virus; and (c) inhibiting the interaction of an agent with a host cell receptor. In addition, eucaryotic cells infected with, and pharmaceutical compositions containing such a recombinant retrovirus are disclosed. Various methods for producing recombinant retroviruses having unique characteristics, and methods for producing transgenic packaging animals or insects are also disclosed.

Abstract: Methods for preserving an infectious recombinant virus for subsequent reconstitution are provided. Within one aspect, the method comprises the steps of (a) combining an infectious recombinant virus with an aqueous solution comprising a saccharide, a high molecular weight structural additive, a buffering component and water to form an aqueous suspension, thereby stabilizing the infectious virus; (b) cooling the aqueous suspension containing the virus to a temperature below the glass transition state temperature or below the eutectic point temperature of the formulation; and (c) removing water from the cooled aqueous suspension by sublimation to form a lyophilized virus having less than 10% water by weight of the lyophilized virus, the virus being capable of infecting mammalian cells upon reconstitution.

Abstract: The present invention provides compositions and methods for utilizing recombinant alphavirus vectors. Also disclosed are compositions and methods for making and utilizing eukaryotic layered vector initiation systems.

Abstract: Lamboid bacteriophage capable of specifically interacting with and delivering nucleic acid molecules to eukaryotic cells are disclosed. Such bacteriophage-derived gene transfer systems target one or more specific receptors on eukaryotic cells, for instance by incorporating mutant tail fiber proteins or by incorporating known ligands for specific eukaryotic receptors into lambda phage. Also disclosed are methods for identifying and producing modified bacteriophage tail fiber polypeptides capable of specifically interacting with eukaryotic transmembrane proteins. Methods of treating diseases using such gene transfer systems are also disclosed.

Abstract: The present invention provides methods and compositions for inhibiting the production of replication competent virus. The invention comprises nucleic acid cassettes encoding a non-biologically active inhibitory molecule which are incorporated into packaging cells and recombinant vector constructs. Upon recombination between various vector construct contained within the producer cell, a biologically active molecule is produced which kills the cell, thereby inhibiting production of replication competent virus.

Abstract: Recombinant retroviruses carrying a vector construct capable of preventing, inhibiting, stabilizing or reversing infectious, cancerous or auto-immune diseases are disclosed. More specifically, the recombinant retroviruses of the present invention are useful for (a) stimulating a specific immune response to an antigen or a pathogenic antigen; (b) inhibiting a function of a pathogenic agent, such as a virus; and (c) inhibiting the interaction of an agent with a host cell receptor. In addition, eucaryotic cells infected with, and pharmaceutical compositions containing such a recombinant retrovirus are disclosed. Various methods for producing recombinant retroviruses having unique characteristics, and methods for producing transgenic packaging animals or insects are also disclosed.