Abstract: The present invention provides a mating-based yeast two-hybrid system for determining whether a test polypeptide interacts with another test polypeptide in the presence or absence of one or more test compounds. The system is useful in detecting protein-protein interactions and in identifying compounds capable of modulating protein-protein interactions.

Abstract: Disclosed are Hansenula polymorpha mutants useful as host cells through which various proteins can be produced as being intact at high yield and a process for preparing recombinant proteins using the host cells. Using various vectors, Hansenula polymorpha is made to be a mutant which is deprived of methanol assimilating ability and incapable of utilizing methanol as a carbon source. This Hansenula polymorpha mutant is used as a high yield host to produce recombinant proteins without continuous feeding of methanol, with the aid of an expression cassette carrying a promoter capable of inducing the expression at a low concentration of methanol. Further, the mutant is also lacking in carboxypeptidase Y, protease Y and/or carboxypeptidase a activity, so the recombinant protein of interest is not degraded at its carboxyl terminal when being expressed in the cell. Thus, intact recombinant protein can be obtained.

Abstract: The present invention concerns a method of genetic modification of a TGB-3 wild type viral sequence for reducing or suppressing the possible deleterious effects of the agronomic properties of a transformed plant or plant cell by the TGB-3 viral sequence, comprising the following successive steps: submitting the sequence to point mutation(s) which allow the substitution of at least one amino-acid into a different amino-acid, selecting genetically modified TGB-3 wild type viral sequences having the point mutation(s) and which are not able to promote cell-to-cell movement of a mutant virus having a dysfunctional TGB-3 wild type viral sequence, when expressed in trans from a replicon, further selecting among the genetically modified TGB-3 viral sequences, the specifically genetically modified sequence which inhibits infection with a co-inoculated wild type virus when the mutant form was expressed from a replicon, and recovering the specifically genetically modified TGB-3 viral sequence.

Abstract: The present invention provides a high-throughput method for identifying a polynucleotide which encodes a transcription factor for controlling the expression of one or more genes in a pathway. In particular, the method is useful for identifying a transcription factor for controlling a gene in a biosynthetic pathway. The invention further provides polynucleotides encoding such transcription factors for controlling the expression of a gene in a biosynthetic pathway, transgenic cells expressing at least one such polynucleotide, and methods for isolating metabolites from such cells or plants.

Abstract: A method is disclosed herein for monitoring the efficiency of an endonuclease digestion using a plasmid specifically designed for that purpose. The plasmid of the present invention comprises at least two polylinker regions containing a plurality of unique restriction sites distributed so that digestion of the plasmid with any two restriction endonucleases whose sites are represented on the plasmid results in two fragments that are sufficiently different in size from the intact plasmid so as to be readily distinguishable therefrom. To ensure this size differential, the polylinker regions of the plasmid are separated by a spacer segment comprising a restriction site-free nucleic acid sequence that is at least about 15% of the length of the intact plasmid.

Abstract: A method for detecting protein-protein interactions is provided, in which two fusion proteins are prepared and allowed to interact with each other. The interaction between the two fusion proteins leads to protein trans-splicing, generating an active and detectable reporter.

Abstract: The present invention relates to a retroviral vector which is especially applicable as a safe gene transfer vehicle for targeted gene therapy. Said retroviral vector comprises one or more promoters inserted in antisense orientation within the 5′ LTR region and one or more coding sequences inserted in antisense orientation within the 3′ LTR region. Both, the promoter as well as the coding sequence, are additionally inserted in such a way as to ensure that the promoter and the coding sequence become duplicated during the process of reverse transcription in a target cell and thus appear in the 3′ as well as in the 5′ LTR region of the resulting provirus in a fashion where the promoter is located upstream of the coding sequence allowing it to drive gene expression. This system avoids any leakiness of gene expression in the packaging cells, and allows expression of transferred genes in the target cell without the necessity for external stimuli.

Abstract: The present invention provides methods and compositions, including kits, for the isolation and purification of mRNA, particularly poly(A) RNA. It concerns the use of isostabilizing salts such as TMAC and TEAC to reduce rRNA carryover during the purification process, thus facilitating the isolation of poly(A) RNA.

Abstract: The present invention solves the problem of integrating multiple copies of a gene of interest by homologous recombination into well defined positions adjacent to conditionally essential genes in a bacterial host strain chromosome, which already comprises at least one copy of the gene of interest in a different position.

Abstract: An assay for the detection of substances agonistic or antagonistic to the mevalonate pathway are disclosed.

Abstract: A Candida albicans gene, CaNik1, is involved in phenotypic switching which is significant because of a direct correlation between the switching and the level of virulence of the organism. A method of screening for anti-fungal pharmaceutical candidates entails bringing a test substance into contact with cells containing a CaNik1 gene or a variant thereof and then monitoring the effect, if any, on the level of expression of the gene.

Abstract: The present invention relates to yeast cells containing the SRB1/PSA1 gene and/or the PKC1 gene or functional derivatives thereof operatively linked to a heterologous inducible promoter and also to methods of regulating yeast cell lysis and flocculation and methods of fermentation using such yeast cells.

Abstract: The present invention relates to receptors for heat shock proteins (HSPs), such as gp96, Hsp70 and Hsp90. The heat shock receptor is associated with the cell membranes of a subset of antigen presenting cells, such as macrophages and dendritic cells. The present invention relates to the use of the heat shock protein receptor positive cells, heat shock protein receptor protein, and heat shock protein receptor genes in methods for screening a molecule for the ability to modulate heat shock protein levels or activities.

Abstract: A 16S rRNA profile derived from Dehalococcoides ethenogenes has been identified and isolated. The profile contains several nucleic acid fragments that are linked to dechlorinating activity. These sequences are set forth in SEQ ID NO:1, SEQ ID NO:8, SEQ ID NO:30 and SEQ ID NO:34.

Abstract: An animal, e.g., transgenic mouse, in which the MSH4 gene is misexpressed. The animal is useful for screening treatments for a number of conditions. Methods for identifying contraceptive agents are also described.

Abstract: Method and compositions for regulating cell cycle progression are disclosed. Compositions include nucleic acids comprising a human Cdc5 gene, antisense gene and fragments thereof and a human Cdc5 protein and polypeptide fragments thereof polypeptide. The consensus DNA binding site for hCdc5 has been described.

Abstract: A method for detecting interactions between first and second interacting molecules at variable sensitivity. This variable sensitivity may be obtained by providing for the overexpression of either a bait hybrid protein containing a DNA binding domain (desensitization) or a prey hybrid protein containing the DNA activation domain for a reporter gene (enhanced sensitivity). The use of exogenous activators of one or the other according to the needs of a particular system is readily accomplished.

Abstract: The present invention relates to a recombinant non-yeast DNA, which encodes a protein of interest, wherein an unmodified DNA corresponding to the recombinant non-yeast DNA contains a region having a high content of codons that are poorly suited to yeasts, wherein a number of the codons that are poorly suited to yeasts are replaced in said region of the recombinant non-yeast DNA with synonymous codons coding for the same amino acid that are well-suited to yeasts, and wherein the number of replaced codons is sufficient to permit expression in yeasts. The present invention also relates to DNA sequences which originate from dicotyledonous or monocotyledonous plants, and in particular plants of the graminae family which are selected from among wheat, barley, oats, rice, maize, sorghum and cane sugar, as well as to vectors and transformed yeasts which contain the DNA sequences of the invention.

Abstract: The present invention provides methods and compositions that enable the experimental determination as to whether any gene in the genome of a diploid pathogenic organism is essential, and whether it is required for virulence or pathogenicity. The methods involve the construction of genetic mutants in which one allele of a specific gene is inactivated while the other allele of the gene is placed under conditional expression. The identification of essential genes and those genes critical to the development of virulent infections, provides a basis for the development of screens for new drugs against such pathogenic organisms. The present invention further provides Candida albicans genes that are demonstrated to be essential and are potential targets for drug screening. The nucleotide sequence of the target genes can be used for various drug discovery purposes, such as expression of the recombinant protein, hybridization assay and construction of nucleic acid arrays.

Abstract: The present invention is directed to translational regulatory elements that mediate the amount of protein produced within a host capable of expressing a construct comprising one or more translational regulatory elements in operative association with a gene of interest. These translational regulatory elements were derived from T1275 (tCUP) and exhibit a high degree of similarity with members of the RENT family of repetitive elements. Translational regulatory elements are disclosed that either increase or decrease he amount of protein produced within the host organism. These translational elements are operative in a wide range of hosts including plant, animals, yeast, fungi and bacteria. Analogs, derivatives and fragments of these translational elements are also disclosed.