Abstract: Method and compositions are provided for introducing alien DNA into a nucleic acid in vivo by providing a plasmid capable of replication in a first host, which is capable of conjugation with a second host into which the alien DNA is to be introduced. The plasmid is characterized by being capable of maintenance in said first host, being incapable of maintenance in said second host, having a region of homology with DNA present in said second host, not self-transmissable but capable of mobilization by a helper plasmid and having a marker, which may be the alien DNA, for selection in said second host. The invention finds particular application for introducing alien DNA into the Ti plasmid of Agrobacterium tumefaciens for introduction into plant cells.

Abstract: Polypeptides, fragments thereof, antisera and monoclonal antibodies thereto are provided for the detection of retroviruses in the HTLV family capable of inducing transformation in human lymphocytes and other immune cells, regulation of viral replication and transformation or infection of human cells.

Abstract: A novel Type II restriction endonuclease isolated from Frankia species DDB13250120 is provided. The enzyme, designated FseI, recognizes the octanucleotide sequence ##STR1## and cleaves Adenovirus 2 DNA at the sites indicated by the arrows.

Abstract: Novel methods and compositions are provided for preventing secondary cataracts. A cytotoxic agent is employed which is introduced into the anterior chamber of the eye to inhibit proliferation of remnant lens epithelial cells after extracapsular cataract extraction. Desirably a non-cytotoxic agent cross-reactive with the cytotoxic agent is introduced prior to introduction of the cytotoxic agent. The agents can be provided as kits.

Abstract: Novel constructs in plasmids are provided for evaluating the efficiency of expression and secretion of structural genes. The constructs provide for transcriptional and translational regulatory regions, a signal sequence and structural gene, which also may be readily excised and substituted, so as to allow for mixing and matching of regulatory regions, signal sequences and genes to evaluate regions for use in the expression of a desired peptide. Particularly, synthetic regions are provided which may be used with other synthetic regions or wild-type regions.Plasmids are provided for screening Bacillus genomic sequences for regulatory regions, particularly promoters employing a structural gene secreting an enzyme which can produce a product which allows for visual detection.

Abstract: Novel oligopeptides are provided having serologic activity for the a determinant of hepatitis B virus surface antigen. Included in the oligopeptide chain are a sequence of at least two cysteines and a lysine in proximity to the cysteines. The oligopeptides can find use in immunoassays, the formulation of vaccines, and the production of antisera.

Abstract: DNA constructs that are useful for providing secretory expression of heterologous polypeptides in yeast comprising a DNA sequence that includes a Kluyveromyces .alpha.-factor leader sequence linked to the heterologous polypeptide by a yeast processing signal. Constructs employing the K. lactis .alpha.-factor leader and processing signal sequences, with and without spacer, linked to prochymosin are exemplified.

Abstract: Phosphatides are shown to have antiviral activity, where the phosphatides are characterized by having a polyunsaturated fatty acid at the sn2 position. By contacting cells susceptible to or infected with a virus, particularly a retrovirus, the proliferation of the virus is substantially inhibited.

Abstract: A novel hybridization diagnostic technique is involved employing micromanipulation of cells for detection of DNA or RNA in the cells. Cellular suspensions are employed, the DNA or RNA purified by microextraction using liquid media having differential solubilization properties, whereby the nucleic acid is retained in an aqueous microdrop. The nucleic acid in the microdrop may then be assayed with a nucleic acid probe in accordance with conventional ways.

Abstract: Recombinantly produced serum albumin is purified in a series of steps, initially employing an alkaline precipitation, optionally followed by incubation with an anion exchange absorbent, followed by affinity chromatography employing a hydrophobic solid phase and using a water-soluble lipid anion as desorbens in the aqueous phase.

Abstract: Novel constructs are provided for expression of physiologically active mammalian proteins in plant cells, either in culture or under cultivation. The constructs provide a promoter functional in a plant host, a structural gene coding for mammalian protein and a terminator functional in a plant host. The construct is introduced into a plant cell to become integrated into the plant genome for expression in the plant cells or plants. The plant cells may be harvested and the mammalian protein isolated in physiologically active form.

Abstract: Serum samples for vitamin B.sub.12 analysis can be pretreated using a combination of peroxy acid and dithiothreitol. The pretreatment makes vitamin B.sub.12 bound to serum binding proteins available for analysis by any of a variety of currently available assay techniques. The pretreatment finds particular use in an assay using solid phase intrinsic factor as a specific binding protein.

Abstract: Kluyveromyces hosts and DNA expression cassettes for use in Kluyveromyces are provided for transcription of endogenous and/or exogenous DNA, and production of peptides, for enhancing production of an endogenous product, or producing an exogenous product. The Kluyveromyces hosts find particular use for secretion of a desired peptide product, where signal sequences may be native to the peptide or provided from endogenous or exogenous signal sequences, including synthetic sequences, functional in Kluyveromyces. A transformation procedure is provided for efficiently transforming Kluyveromyces.

Abstract: Fruit specific regulatory regions are identified employing cDNA screening. The resulting fruit specific regulatory regions are manipulated for use with foreign sequences for introduction into plant cells to provide transformed plants having fruit with a modified phenotypic property. The invention is exemplified with a tomato fruit specific promoter which is active throughout the stages of fruit ripening.

Abstract: Novel methods and compositions are provided for preventing secondary cataracts. A cytotoxic agent is employed which is introduced into the anterior chamber of the eye to inhibit proliferation of remnant lens epithelial cells after extracapsular cataract extraction. Desirably a non-cytotoxic agent cross-reactive with the cytotoxic agent is introduced prior to introduction of the cytotoxic agent. The agents can be provided as kits.

Abstract: Novel compositions and methods employing the compositions are provided involving eukaryotic plasmids having a replication system or replicon from an extrachromosomal element, a chromosomal replicator, and the regulatory signals of the structural gene for yeast 3-phosphoglycerokinase controlling expression of a structural gene. The plasmids find particular use in yeast for production of poly(amino acids).

Abstract: Plant host acceptable microorganisms, which are ice nucleation deficient and use at least one nutrient from the plant also used by ice nucleating native microorganisms, are applied to a plant part at an early stage in the growth cycle. The multiplication of the native ice nucleating microorganisms is inhibited, so that under normal frost conditions encountered in the field, frost damage is substantially diminished. The non-nucleating microorganisms may be obtained by special selection procedures, selecting from naturally occuring microorganisms or mutagenized microorganisms, where additionally the organisms may be transformed to provide for other desirable properties.

Abstract: Live vaccines are provided and methods for preparing the live vaccines for protection of a host from a pathogenic microorganism. The vaccines are prepared by introducing at least one modification in a gene involved in at least one, normally at least two, biosynthetic pathways, involving the production of products which are unlikely to be found in the disease susceptible host. The modification results in a gene change which cannot be repaired by a single step e.g. polynucleotide deletions and inversions. Where the aro gene suffers such a change, the resultant auxotrophic mutants require aromatic amino acids, p-aminobenzoic acid and 2,3-dihydroxybenzoic acid or a highly concentrated source of absorbable iron. The auxotrophic mutations have substantially reduced or nonexistent virulence, while retaining the desired immunogenicity to initiate the immunogenic response. Various techniques can be employed for providing the desired change.Salmonella typhimurium strain SL1479 was deposited at the ATCC on Sept.

Abstract: Novel polypeptides, polynucleotide sequences, DNA constructs and compositions are provided for the preparation and use of polypeptides associated with naturally occurring polypeptides found in brains. The small molecular weight polypeptides are growth inhibitors for neoplastic cells without inhibiting normal cells. The polypeptides comprise specific regions which are highly conserved, separated by less conserved regions. The polypeptides find use in inhibiting neoplastic growth and detecting receptors for the polypeptides. Antibodies are provided in conjunction with the polypeptides, which may be used together or separately for detecting the presence of the neoplastic cell retarding polypeptides.