The invention provides DNA construct comprising in the 5' to 3' direction of transcription a promotor functional in a plant, an intron, a nucleotide sequence of interest, and a terminator region, said DNA construct comprising two non-identical recombination sites, where one of said non-identical recombination site is contained within said intron.

The present invention provides methods and compositions for the efficient and enhanced secretion of a protein of interest from a host cell. In specific, proteins are secreted through the Sec-dependent pathway, involving the spoIIIJ and/or yqjG gene product(s). In some embodiments, expression of the spoIIIJ and/or yqjG gene product(s) is modulated by a promoter operably linked to the gene.

The present invention provides methods for building DNA constructs in vitro, transforming such constructs into competent Bacillus strains with good efficiency, and generating populations of mutants. Also provided is a method to assemble DNA constructs in situ.

The present invention provides methods for building DNA constructs in vitro, transforming such constructs into competent Bacillus strains with good efficiency, and generating populations of mutants. Also provided is a method to assemble DNA constructs in situ.

The present invention provides a method for identifying species and subspecies in a biological sample through the selective amplification of segments of nucleic acid that code a target region of the cytoplasmatic beta-actin protein, which is present in all the organisms concerned. The method comprises DNA extraction from the sample; amplification of divergent segments of the cytoplasmatic beta-actin gene by PCR or an equivalent technique, using primers of regions with high evolutionary conservation between species and subspecies; and identification of the amplified segment by comparison of its size in base pairs with a pre-established standard of sizes and/or identification of the amplified segment by DNA sequencing and comparison of the resulting sequence with the specific sequence of each species or subspecies present on a computer database.

Novel vectors are described that incorporate, inter alia, a novel 3' gene trap cassette which can be used to efficiently trap and identify previously unknown cellular genes. Vectors incorporating the described 3' gene trap cassette find particular application in gene discovery and in the production of mutated cells and animals.

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.

The present invention generally relates to non-human transgenic animals expressing human cellular markers, including CD20 and/or preferably CD16.

A method is provided for increasing the specific activity of a type III antifreeze protein when said protein is prepared by expression in a heterologous fungal species of a gene encoding the protein sequence, by means of reducing the extent of glycosylation of the protein.

The invention relates to the growth temperature of organisms, especially plants and microorganisms and the manipulation of the tolerable cultivation temperature. More specifically, the present invention relates to the expression of heterologous proteins in microorganisms, and especially to the heterologous expression of heat sensitive proteins in bacteria, either gram-negative or gram-positive. In a first aspect, the present invention provides a method for manipulation of cells and the resultant cells, wherein at least one gene from a psychrophilic micro organism coding for at least one chaperone or chaperonin is expressed. Such cells are selected among cultivated eukaryotic cells, i.e. animal and plant cells and entire plants, gram-negative and gram-positive bacteria, fungi and yeasts.