Abstract: The invention represents the disclosure of a novel insect intestinal mucin comprising two nearly identical isoforms, IIM14 and IIM22 respectively. These isoforms of the IIM protein have been identified and cloned using T. ni larva. The cDNA and amino acid sequences have been determined and are disclosed. Both IIM isoforms have an approximate molecular mass of 400 kDa. These sequences once disclosed are useful for the production of transgenic or recombinant vectors including viral, microorganism cell, plant, or animals, wherein the virus, microorganism, cell, plant, or animal is the product of an insertion of a gene expression vector including a DNA that encodes an IIM protein sequence. Thereafter the engineered host of the IIM DNA sequence is capable of expressing said IIM protein in a functional form. Also useful is a purified and isolated recombinant DNA sequence comprising a DNA sequence that codes for an IIM protein.

Abstract: A method of infecting insects is disclosed; the method utilizes a form of a baculovirus which is highly efficient at establishing infection and is normally destined to become occluded within the polyhedrin or granulin--Pre-occluded Virus (POV). Specifically, the POV as derived from a polyhedrin-minus or granulin-minus (lacking a functional polyhedrin or granulin gene) baculovirus is fed to insect larvae per os resulting in high infection rates. The stabilization and use of the POV form of polyhedrin-minus baculoviruses for recombinant protein production and as an insecticide is also disclosed.

Abstract: We have studied the regulation of the extracellular chymoelastase protease (Pr1) of Metarhizium anisopliae, an enzyme involved in the penetration of insect cuticle by Metarhizium and other entomopathogenic fungi. We report here the isolation and characterization of a Pr1 cDNA clone with a full length insert. Pr1 is synthesized as a large precursor (40.3 kDa) containing a signal peptide and a propeptide and the mature protein is predicted to have a relative molecular mass of 28.6 kDa. The primary structure of Pr1 shares extensive homology (30-60%) with enzymes of the subtilisin subclass of the serine endopeptidases and the serine, histidine and aspartyl components of the active site in subtilisins are preserved. The genes coding for chymoelastase or slightly altered versions thereof, can be used to transform various organisms (i.e. fungi, viruses, plants, bacteria, etc.) such that the transformed organisms are capable of producing chymoelastase in recoverable quantities.

Abstract: Disclosed are methods of increasing exogenous protein expression in a cell or a transgenic plant. Constructs, i.e., vectors, DNA fusions and polynucleotides, for use in conjunction with the methods to cause increased exogenous protein expression are also disclosed. These constructs generally include intron 1 and/or intron 2 of the PAT1 gene. Additionally disclosed are cells, including recombinant cells, and plant lines transformed with the described constructs. In particular, a cultivated, transgenic food plant, the genome of which has been augmented through the genomic introduction of a preselected exogenous protein gene not found in the genome of non-transformed parentage of the plant is described. Also described are seed, progeny and cells of the described transgenic food plant.

Abstract: Disclosed herein is a novel baculovirus cloning system. The new cloning system is a marker-rescue system, using an essential gene, e.g. gp64. In this system, a gene essential for viral replication, growth, or propagation in cell culture is removed from or inactivated in the viral genome. Once a null baculovirus is created, it is propagated in a host cell that expresses the essential protein or a functional homolog. For cloning into the baculovirus containing the null-mutation, the virus is used to infect wild type host cells and the same cells are transfected with a plasmid that contains the essential gene, or a functional homolog, linked to a foreign gene under the control of a selected promoter. The baculovirus is "rescued" by the rescue gene linked to the foreign gene and is able to propagate normally and express the foreign gene.

Abstract: This disclosure relates to an isolated and cloned DNA from a granulovirus virus which comprises an amino acid sequence of the vital gene encoding a polypeptide isolated from occlusion bodies of certain baculoviruses and which polypeptide possesses the biological activity of enhancing baculovirus infectivity. Such proteins termed herein as "enhancins" are found within the viral occlusion body, have a disruptive effect on the insect peritrophic membrane (PM) proteins, and/or interact with the midgut epithelium in such a manner as to permit the increased adsorption, penetration and uptake of virus particles by midgut cells with a concomitant increase in host mortality. Disclosed herein is a recombinant DNA sequence which codes for the enhancin protein of the Helicoverpa armigera granulovirus virus. The DNA sequence is shown in SEQ. ID. NO.: 1 and the open reading frame is shown in SEQ. ID. NO.: 1: base pairs 271-2976. The amino acid sequence of the enhancin protein is shown in SEQ. ID. NO.: 2.

Abstract: This disclosure presents the establishment of a new cell line from Pseudaletia unipuncta embryos. This cell line demonstrated the ability to produce high numbers of baculoviruses in cell culture. These virus particles are found internally in the cells in occlusion bodies. In the study of Pseudaletia unipuncta two baculoviruses were found to infect this species: P. unipuncta nuclear polyhedrosis virus (PuNPV), and P. unipuncta granulosis virus (PuGV). In addition, the cell line was also selected and cultured for its ability to grow in suspension while maintaining high levels of OB production.

Abstract: A method of infecting insects is disclosed; the method utilizes a form of a baculovirus which is highly efficient at establishing infection and is normally destined to become occluded within the polyhedrin or granulin--Pre-occluded Virus (POV). Specifically, the POV as derived from a polyhedrin-minus or granulin-minus (lacking a functional polyhedrin or granulin gene) baculovirus is fed to insect larvae per os resulting in high infection rates. The stabilization and use of the POV form of polyhedrin-minus baculoviruses as an insecticide is also disclosed.

Abstract: Method and apparatus for rearing insects is disclosed. Information concerning the physical and dietary needs of the insect as well as behavioral characteristics are utilized to maximize the number of larvae reared per unit surface area of diet and per unit of rearing area while minimizing the amount of labor and materials required. An enclosed rearing unit is provided which can be located within an appropriate environment for rearing the insects. There are three sections within the rearing unit: 1) a diet space, 2) a larval space, and 3) a frass space. The diet space includes an appropriate diet medium for the insects. The larval space is located below the diet space and includes a series of vertical partitions perpendicular to and in contact with or nearly in contact with the diet medium such that the insect larvae are able to disperse themselves over the partitions.

Abstract: Normally anchorage-dependent insect cell lines are adapted to replicate under suspension conditions by addition of heparin to the culture medium and selection for resulting suspension-tolerant cells.

Abstract: An insect cell line has been established and characterized, derived from embryonic tissue (BTI-TN-5B1-4, ATCC CRL 10859) of Trichoplusia ni (cabbage looper). The line is susceptible to various baculoviruses, including TnSNPV and AcMNPV, and may be used to replicate such viruses for use as insecticides or otherwise.

Abstract: Two new insect cell lines have been established and characterized; the cell lines were derived from midgut (BTI-TN-MG1, ATC CRL 10860), and embryonic tissue (BTI-TN-5B1-4, ATC CRL 10859) of Trichoplusia ni (cabbage looper). The lines are susceptible to various baculoviruses, including TnSNPV and AcMNPV, and may be used to replicate such viruses for use as insecticides or otherwise.

Abstract: A method is provided for protection of biological materials from the stresses of air-drying and also from destructive reactions, such as oxidation and free-radical attack, which degrade the materials during long-term storage. The method involves drying the materials, which may contain potentially destructive agents such as free-radical generators or reducing sugars, in the presence of a vitrifying substance, and under conditions which allow the protective substance to become vitrified.

Abstract: A host microorganism is genetically and stably modified by the insertion into any of its non-essential chromosomal location of a non-homologous, recombinant foreign DNA fragment, maintaining an insertion of a luxAB gene of a selected bioluminescent bacterium such as V. harveyi, such that the expression of the luxAB genes causes the production of a luciferase enzyme which, in turn, catalyzes a light-emitting reaction in the presence of the appropriate substrate. X-ray film can be used to quantify the light being emitted from a microorganism through the use of plural droplets containing the same microorganism, each with a known and related cell (or plasmid) count.

Abstract: A method is provided for protection of biological materials from the stresses of air-drying and also from destructive reactions, such as oxidation and free-radical attack, which degrade the materials during long-term storage. The method involves drying the materials, which may contain potentially destructive agents such as free-radical generators or reducing sugars, in the presence of a vitrifying substance, and under conditions which allow the protective substance to become vitrified.

Abstract: Nuclear polyhedrosis viruses, for example, Autographa californica nuclear polyhedrosis virus (AcMNPV), useful in the control of lepidopterous larvae such as the larvae of the cabbage looper Trichoplusia ni, have been found to have enhanced infectivity when mixed with certain proteins obtained from the granulin fraction of Trichoplusia ni granulosis virus (TnGV) or Heliothis armigera granulosis virus (HaGV), and from the polyhedrin fraction of AcMNPV viruses. The proteins from the TnGV granulin fraction have molecular weights of about 101 and about 104 Kd. The enhanced infectivity is correlated to biochemical and structural changes in the T.ni peritrophic membrane.

Abstract: Nuclear polyhedrosis viruses, for example, Autographa californica nuclear polyhedrosis virus (AcMNPV), useful in the control of lepidopterous larvae such as the larvae of the cabbage looper Trichoplusia ni, have been found to have enhanced infectivity when mixed with certain proteins obtained from the granulin fraction of Trichoplusia ni granulosis virus (TnGV) or Heliothis armigera granulosis virus (HaGV), and from the polyhedrin fraction of AcMNPV viruses. The proteins from the TnGV granulin fraction have molecular weights of about 101 and about 104 kd. The enhanced infectivity is correlated to biochemical and structural changes in the T.ni peritrophic membrane.

Abstract: A recombinant subunit vaccine for protecting dogs against infection caused by canine parvovirus comprising VP-2 protein produced during replication of a recombinant baculovirus in insect tissue culture cells or insects which are a permissive host for the replication of selected baculoviruses.

Abstract: A procaryotic microorganism and a method for its production is provided wherein the microorganism contains at least one stable foreign DNA portion in the chromosome. The disclosed microorganisms and their progeny are substantially free of genetic rearrangement involving the foreign DNA. In a preferred embodiment, cyanobacteria are employed. The microorganisms are produced by introducing into the cell an insertion vehicle that contains foreign DNA ligated between two portions of DNA homologous to adjacent portions of the recipient's chromosome.