Abstract: Phycotoxins are purified from a mixture of phycotoxins produced in a continuous process. Cyanobacteria are produced in a continuous culture, then lyzed, the cells pelleted and extracted, and the extract purified using an organic solvent-aqueous mixture and repeated passage through a diatomaceous earth column. The column is washed with acetic acid, then the neosaxitoxin extracted with an alcohol-water mixture. The eluate is passed through activated charcoal columns, which are washed with distilled water to remove the retained pigments and impurities, the further purified by HPLC. In one embodiment, the process produces only neosaxitoxin and saxitoxin. In another embodiment, the process produces only GTX2/3.

Abstract: Methods for producing phycotoxins from natural sources, wherein the phycotoxins have a definite compositional profile are described herein. In one embodiment, the phycotoxins are produced by cyanobacteria. In one embodiment, the phycotoxins are produced by continuously culturing cyanobacteria under strictly controlled conditions in order to produce a definite compositional profile. In another embodiment, organic nutrients are added to the culture that allows for higher concentrations of neosaxitoxin and saxitoxin or gonyaulatoxins 2 and 3 per weight of the algae. The phycotoxins are isolated primarily from the bacteria but can also be isolated from the culture medium. In one embodiment, the cyanobacteria produce only neosaxitoxin and saxitoxin in a ratio of about 6:1, 5:1, 4:1, or 3:1. In a preferred embodiment, the amount of saxitoxin is less than 20% by weight of the total amount of neosaxitoxin and saxitoxin produced. In another embodiment, the cyanobacteria produce only GTX2 and GTX 3.

Abstract: Phycotoxins are purified from a mixture of phycotoxins produced in a continuous process. Cyanobacteria are produced in a continuous culture, then lyzed, the cells pelleted and extracted, and the extract purified using an organic solvent-aqueous mixture and repeated passage through a diatomaceous earth column. The column is washed with acetic acid, then the neosaxitoxin extracted with an alcohol-water mixture. The eluate is passed through activated charcoal columns, which are washed with distilled water to remove the retained pigments and impurities, the further purified by HPLC. In one embodiment, the process produces only neosaxitoxin and saxitoxin. In another embodiment, the process produces only GTX2/3.

Abstract: Phycotoxins are purified from a mixture of phycotoxins produced in a continuous process. Cyanobacteria are produced in a continuous culture, then lyzed, the cells pelleted and extracted, and the extract purified using an organic solvent-aqueous mixture and repeated passage through a diatomaceous earth column. The column is washed with acetic acid, then the neosaxitoxin extracted with an alcohol-water mixture. The eluate is passed through activated charcoal columns, which are washed with distilled water to remove the retained pigments and impurities, the further purified by HPLC. In one embodiment, the process produces only neosaxitoxin and saxitoxin. In another embodiment, the process produces only GTX2/3.

Abstract: The invention concerns a method for detecting a known function, from nucleic acids present in a sample, characterized in that it comprises the following steps: (a) preparing, from the nucleic acids of the sample, nucleic acid molecules comprising the gene(s) coding for the protein(s) corresponding to said function, and the control elements required for the transcription and the translation of said gene(s); (b) in vitro transcription and translation of the nucleic acid molecule prepared in step (a); (c) detecting and/or measuring the function corresponding to the protein(s) produced in step (b).

Abstract: The invention relates to methods of determining the mutational load of a gene bank obtained by random mutagenesis of a gene of interest by preparing a chart linking the mutational load of a gene bank obtained by random mutagenesis of a model gene with the fraction of mutated model genes observed in the bank; performing, in parallel, random mutagenesis of the model gene used for preparing the chart and the gene of interest to obtain the corresponding mutated gene banks; determining the mutational load of the gene bank obtained using the model gene on the basis of the chart plotted; and applying a correction factor to the mutational load of the mutated model gene bank to determine the mutational load in the gene bank of the mutated genes of interest.

Abstract: The present invention relates to a newly identified hydrolase from thermophilic microorganisms having thermostable properties, and more specifically, to a novel thermostable hydrolase showing high activity at high temperatures.

Abstract: The present invention relates to a method used to determine the mutational load of a gene bank obtained by means of random mutagenesis of a gene of interest comprising: a) the preparation of a chart linking the mutational load (ML) of gene bank obtained by means of random mutagenesis of a model gene with the fraction of mutated model genes observed in said bank; b) the random mutagenesis of the model gene used for the preparation of the chart for step (a) and the gene of interest to obtain the corresponding mutated gene banks; c) the determination of the mutational load (ML) of the gene bank obtained using the model gene in step (b) on the basis of the chart plotted in step (a); d) the application of a correction factor (CF) to the mutational load (ML) of the mutated model gene bank determined in step (c) to determine the mutational load (ML) of the banks of mutated genes of interest from step (b).

Abstract: The invention relates to a fragmentation process that depends on mismatches between two strands of parental polynucleotides. One embodiment, comprises a method for preparing polynucleotide fragments of DNA comprising formation of heteroduplex molecules by hybridizing polynucleotides. The invention also provides a method and process of forming fragments which can be used with any shuffling process or combination of shuffling processes.

Abstract: The present invention provides methods and test-kits for rapidly detecting and/or quantifying a group of organism, cultivable or not cultivable, sharing the same enzymatic activity. In particular, the methods and kits can be used for example in the field of cosmetics, where the activities of (microbial) enzymes, which are responsible for malodor or dandruff, are detected as an indicator for the presence of particular groups of microorganisms. Furthermore, the effect on substances acting on the causative microbes can be determined rapidly, easily and at low cost.

Abstract: The object of this invention is a process for the creation of at least one recombinant polynucleotide sequence comprising a step of oriented ligation of fragments derived from a bank of at least two polynucleotide sequences, and optionally cloning the recombinant polynucleotide sequences, and the selection of polynucleotide sequences offering advantageous characteristics compared to one or several reference sequences.

Abstract: Ligation-mediated method of recombining polynucleotides in vitro. Polynucleotides from a library are fragmented and the fragments are hybridized to an assembly template. The hybridized fragments are iteratively re-hybridized and ligated until the ends of the hybridized fragments are adjacent to the ends of other hybridized fragments on the assembly template. A final ligation produces recombined polynucleotides.

Abstract: Ligation-mediated method of recombining polynucleotides in vitro. Polynucleotides from a library are fragmented and the fragments are hybridized to an assembly template. The hybridized fragments are iteratively re-hybridized and ligated until the ends of the hybridized fragments are adjacent to the ends of other hybridized fragments on the assembly template. A final ligation produces recombined polynucleotides.

Abstract: The present invention relates to a method of detection in vitro of a target substance in a sample. The sample can comprise, among other things, a nucleic sequence or more generally any type of substance. In particular, the present invention provides a method of detection comprising (i) specificly labeling a substance with a reporter gene and any sequences necessary for the in vitro expression of the reporter gene; (ii) in vitro transcription and translation of the reporter gene; and (iii) in vitro detection of a reporter protein coded by the reporter gene.

Abstract: The invention concerns a method for determining the activity of a substance using a functional test, characterized in that it consists in detecting and/or measuring the variation of a known function corresponding to one or several proteins produced in vitro in the presence and in the absence of said substances or to the substance in the presence or in the absence of one or several proteins produces in vitro.

Abstract: The invention relates to a fragmentation process that depends on mismatches between two strands of parental polynucleotides. One embodiment, comprises a method for preparing polynucleotide fragments of DNA comprising formation of heteroduplex molecules by hybridizing polynucleotides. The invention also provides a method and process of forming fragments which can be used with any shuffling process or combination of shuffling processes.

Abstract: Method and kit for using in vitro expression to discover nucleic acids that encode desired functions. Either the existence, presence, identity, properties or function of one or more of the nucleic acids from the sample is unknown to at least the experimenter performing the method or using the kit.

Abstract: Method of gene shuffling using oriented ligation, whereby at least two fragments are adjacently hybridized on an assembly template. Invention is particularly aimed at generating novel polynucleotides that differ in some advantageous respect compared to a reference sequence. Invention further includes sequences created by the method, hosts and vectors containing same, and proteins translated therefrom.

Abstract: The present invention is directed to a process for the separation and characterization of the functions potentially present in a biological sample containing nucleic acids, characterized in that it comprises the steps of preparation of nucleic acid fragments starting from said sample, association of each one of said fragments with a vector molecule, isolation of each fragment associated with a vector molecule or with a part of each construction composed of a fragment associated with a vector molecule, in vitro treatment of each fragment associated with a vector molecule or of a part of each construction composed of a fragment associated with a vector molecule to obtain transcripts, and testing of the function of the transcripts or of the proteins encoded thereby after translation of said transcripts.