46 results about "Low copy number" patented technology

This invention provides a series of low-copy number plasmids comprising restriction endonuclease recognition sites useful for cloning at least three different genes or operons, each flanked by a terminator sequence, the plasmids containing variants of glucose isomerase promoters for varying levels of protein expression. The materials and methods are useful for genetic engineering in microorganisms, especially where multiple genetic insertions are sought.

Provided are methods of producing low copy number circularized nucleic acid variants that can be distributed to reaction volumes. The methods include providing a template nucleic acid; producing a population of clonal nucleic acids from the template nucleic acid; generating a set of partially overlapping nucleic acid fragments from the population of clonal nucleic acids; circularizing the partially overlapping nucleic acid fragments to produce circularized nucleic acid variants; and aliquotting the circularized nucleic acid variants into reaction volumes. Related compositions of nucleic acid templates are also provided.

Provided herein are products and processes for the amplification, detection and sequencing of short-stranded nucleic acid in the presence of a high background of long-stranded genomic material (e.g., host or maternal nucleic acids). The methods rely on the use of inside and outside primers introduced at varying concentrations, as well as universal amplification reactions that preferentially amplify short, low copy number nucleic acid.

The invention relates to methods of selecting and maintaining a population of pigs having a low copy number of porcine endogenous retrovirus, and the use of such pigs as a source of cells, tissue and / or organs suitable for xenotransplantation. The invention further relates to methods for selecting cells, tissue and / or organs from such pigs for suitability for use in xenotransplantation.

Compositions, methods and kits for detecting HIV-1 nucleic acids using nucleic acid amplification. Particularly described are oligonucleotides that are useful as hybridization probes and amplification primers for detecting very low levels of HIV-1 nucleic acids by real-time monitoring of amplicon production. The invented assays are characterized by high levels of precision in the quantitation of HIV-1 targets at low copy numbers, and by accurate detection of different HIV-1 subtypes, including M group and O group variants.

The invention relates to a method for quantitatively detecting white spot syndrome virus through a TaqMan probe. The method comprises the steps: firstly, extracting genome DNA of a sample to be detected; then designing a primer and the TaqMan probe; finally, performing fluorogenic quantitative PCR detection on the genome DNA of the sample to be detected through the designed primer and the designedTaqMan probe. The detection method disclosed by the invention has the advantages of high detection flexibility and good specificity, short used time, no false positive situation generation, application to molecular disease detection and early warning of prawn farming in China and good application prospect; furthermore, the lowest copy number which can be detected by the detection method is 7.8.

A process for constructing a catalogued nucleic acid library in which the proportional representation of the constituents is adjusted to advantage through the use of disclosed technologies for positive and negative selection. The resultant benefit is that significantly fewer library constituents will need to be screened in order to identify a potentially desired constituent. Moreover, library constituents that previously would have been essentially “lost” are now recoverable. Preferred embodiments of this invention include the cataloguing, normalization, and enrichment of library constituents. By way of example, but not limitation, this technology is serviceable for constructing a library that contains an adequate representation of desirable constituents that (1) are initially found in low-copy numbers within a sample source or (2) originate from an organism that is problematic to culture. Applicable uses of this invention include any library-screening endeavor previously hindered by logistical impediments.By expanding previous logistical frontiers this invention allows for a novel generation of previously unattainable molecules—particularly molecules that are “unclonable” from conventional, unadjusted libraries—to now be detected, cloned, manipulated, expressed, studied, and used. By disclosing the construction and screening of high yielding nucleic acid libraries from mixed and uncultivated organisms, the instant technology eclipses former boundaries in the area of biological discovery and enables the full breadth of biological diversity to be accessed in the search for previously undiscovered genes and gene products. The benefits of the present invention are seen to extend to areas of diagnosis, medicine, agriculture, manufacturing, and academia.

The invention discloses a pC series plasmid as well as a construction method and application thereof, wherein the pC series plasmid comprises a rep gene, a rop gene, a lacZ gene and an antibiotics resistance gene which are sequentially connected with one another end to end. The pC series plasmid has the characteristics of small molecular weight, large capacity, low copy number, high stability andthe like, can be used for instable genes with complicated structures and repeated sequence, cloning, screening and sequencing long section of genes, genome construction and other gene engineering fields.

The invention discloses a pC series plasmid as well as a construction method and application thereof, wherein the pC series plasmid comprises a rep gene, a rop gene, a lacZ gene and an antibiotics resistance gene which are sequentially connected with one another end to end. The pC series plasmid has the characteristics of small molecular weight, large capacity, low copy number, high stability andthe like, can be used for instable genes with complicated structures and repeated sequence, cloning, screening and sequencing long section of genes, genome construction and other gene engineering fields.

Methods and systems for autoinduction of gene expression, without the need to add exogenous inducers. A dual genetic element system, which includes a first, high copy number genetic element comprisinga first gene of interest that is under the control of an inducible promoter, and a second, low copy number genetic element comprising a gene encoding a transcriptional factor which, upon expression,regulates transcription from the inducible promoter, wherein activation of transcription from the inducible promoter does not require addition of an exogenous inducer.

The invention provides a kit for detection BCR diversity. The kit comprises a set of multiple PCR primers. The multiple PCR primer contains upstream primers and downstream primers. The upstream primers are composed of a group of sequences which have a one-to-one correspondence with nucleotide sequences as shown in SEQ ID NO:1- SEQ ID NO: 13. Sequences in the upstream primer have 0-3 more or less nucleotides than the corresponding sequences in the SEQ ID NO:1- SEQ ID NO:13. The downstream primers are composed of a group of sequences which have a one-to-one correspondence with nucleotide sequences as shown in SEQ ID NO:14- SEQ ID NO:17. Sequences in the downstream primer have 0-3 more or less nucleotides than the corresponding sequences in the SEQ ID NO:14- SEQ ID NO:17. By the use of the kit, BCR sequences of human can be efficiently obtained, human specific BCR CDR3 sequences are obtained, and detection rate of low-copy-number B cell clones can be raised.

The invention relates to methods of selecting and maintaining a population of pigs having a low copy number of porcine endogenous retrovirus, and the use of such pigs as a source of cells, tissue and / or organs suitable for xenotransplantation. The invention further relates to methods for selecting cells, tissue and / or organs from such pigs for suitability for use in xenotransplantation.

This invention provides a method for generating transgenic plants with a low copy number. Plant cells are transformed with polynucleotides containing transcriptional cassettes designed to trigger silencing of a gene which is essential for the plant cell to survive the transformation and regeneration process. The present invention enables the recovery of an increased number of transgenic plants which have only one copy of each desired transcriptional cassette.

Genes are expressed by culturing cells comprising a host chromosome comprising an integrated artificial chromosome comprising recombinant genes, under conditions whereby each recombinant gene is expressed copy number dependently and position independently. Deletions increase expression from recombinant gene(s) inserted into the artificial chromosome.

This invention provides a method for generating transgenic plants with a low copy number. Plant cells are transformed with polynucleotides containing transcriptional cassettes designed to trigger silencing of a gene which is essential for the plant cell to survive the transformation and regeneration process. The present invention enables the recovery of an increased number of transgenic plants which have only one copy of each desired transcriptional cassette.