专利号:WO-9640901-A1 优先权日:1995-06-07 标题 :Template and primer based synthesis of enzymatically cleavable oligonucleotides 发明人:DATTAGUPTA NANIBHUSHAN; KACIAN DANIEL LOUIS; LACKEY DAVID BRUCE 权利人:GEN PROBE INC 摘要:The present invention relates to methods of enzymatically synthesizing chirally pure oligonucleotides. In particular, it relates to methods of enzymatically synthesizing oligonucleotides using optionally digestible templates which are capable of directing the synthesis of an oligonucleotide product which can optionally be cleaved into two or more individual oligonucleotides, optionally using 3'-ribonucleotide primers. In this manner, more than one oligonucleotide having the same or different length and/or sequence can be simultaneously synthesized. The oligonucleotides optionally have at least one phosphorothioate linkage, and contain certain restriction endonuclease recognition sites herein identified as cleavable within phosphorothioate oligonucleotides, thus facilitating the separation and purification of the oligonucleotides after synthesis.
专利号:US-5916777-A 优先权日:1995-06-07 标 题:Enzymatic synthesis of oligonucleotides using 3'-ribonucleotide primers 发明人:KACIAN DANIEL LOUIS; DATTAGUPTA NANIBHUSHAN; LACKEY DAVID BRUCE 权利人:GEN PROBE INC 摘要:The present invention relates to methods of enzymatically synthesizing oligonucleotides. In particular, it relates to the use of 3'-ribonucleotide primers which effectively serve to initiate oligonucleotide synthesis, and can also be easily cleaved from synthesis products and reused in subsequent synthesis reactions.
专利号:EP-1543168-B1 优先权日:2002-09-27 标 题:Method for assaying replication of hbv and testing susceptibility to drugs 发明人:DURANTEL DAVID; DURANTEL SANDRA; TREPO CHRISTIAN; ZOULIM FABIEN 权利人:INST NAT SANTE RECH MED 摘要:Measuring the replication capacity of hepatitis B virus (HBV), e.g. HBV in a biological sample, possibly in the presence of a pharmaceutical product, and particularly an antiviral agent, is new. Measuring the replication capacity of hepatitis B virus (HBV), e.g. HBV present in a biological sample, possibly in the presence of a pharmaceutical product, particularly an antiviral agent, comprises: (a) optional extraction of nucleic acids contained in the sample; (b) PCR amplification of HBV nucleic acids using at least 2 primer pairs to obtain at least 2 amplified HBV genomic fragments representing more-than-full-length HBV genome; (c) cloning the fragments obtained into a vector; (d) transfecting or transducing susceptible cells with the vector; (e) culturing the transfected or transduced cells in conditions allowing synthesis of HBV pregenomic RNA (pgRNA) from the cloned HBV DNA; (f) optionally treating the cultured cells with a pharmaceutical product, particularly an antiviral agent; and (g) determining the replication capacity of the HBV, and the effect of any pharmaceutical product used on viral gene expression and/or viral replication. Independent claims are also included for: (1) a polynucleotide useful as primer for HBV amplification, comprising a sequence selected from 21 sequences of 23-42 bp (SEQ ID NO: 1-21) given in the specification; (2) a primer pair for HBV amplification comprising: (a) a forward primer comprising SEQ ID NO: 1-12, and/or a reverse primer comprising SEQ ID NO: 13 or SEQ ID NO: 14; (b) a forward primer comprising SEQ ID NO: 15 and/or 16 or 19, and a reverse primer comprising SEQ ID NO: 17 and/or SEQ ID NO: 18; (c) a forward primer comprising SEQ ID NO: 19, and a reverse primer comprising SEQ ID NO: 17 and/or SEQ ID NO: 18; or (d) a forward primer comprising SEQ ID NO: 20 or 22, and a reverse primer comprising SEQ ID NO: 21 or 23; (3) kits for HBV amplification, comprising a primer pair of (2); (4) a vector comprising a more-than-full length HBV genome as defined above, and a promoter modified in the 5' by the presence of a restriction site in the 5' of the +1 of transcription, where the +1 of transcription of the more-than-full length HBV genome and of the promoter are fused, and the promoter controls the synthesis of a pgRNA from the more-than-full length HBV genome post-cell-transfection; and (5) a baculovirus or cell line comprising the vector. ACTIVITY : Virucide. MECHANISM OF ACTION : None given.
专利号:US-8008459-B2 优先权日:2001-01-25 标 题 :Concatemers of differentially expressed multiple genes 发明人:GOLDSMITH NEIL; SORENSEN ALEXANDRA M P SANTANA; NIELSEN SOREN V S; NAESBY MICHAEL 权利人:EVOLVA SA 摘要:In the present invention are disclosed concatemers of concatenated expression cassettes and vectors that enable the synthesis of such concatemers. The concatemer comprises in the 5′→3′ direction a cassette of nucleotide sequence of the general formula [rs2-SP-PR-X-TR-SP-rs1]n wherein rs1 and rs2 together denote a functional restriction site, SP individually denotes a spacer of at least two nucleotide bases, PR denotes a promoter, capable of functioning in a cell, X denotes an expressible nucleotide sequence, TR denotes a terminator, and SP individually denotes a spacer of at least two nucleotide bases, and n>/=2, and wherein at least a first cassette is different from a second cassette. The main purpose of these concatemers is the controllable and co-ordinated expression of large numbers of heterologous genes in a single host. Furthermore, the invention relates to a concatemer of cassettes of nucleotide sequences and a method for preparing the concatemers. In a further aspect, the invention relates to transgenic host cells comprising at least one concatemer according to the invention, as well as to a method for preparing the transgenic host cells. Finally, the invention relates to a vector comprising a cassette of nucleotides, a method for preparing said vector, a nucleotide library comprising at least two primary vectors each comprising a cassette of nucleotides, a method for preparing the library.
专利号:US-2006099589-A1 优先权日:2002-07-23 标题:Gene shuffling by template switching 发明人:PEDERSEN HENRIK; THISTED THOMAS; MOLLER HANS T 权利人:NUEVOLUTION AS 摘要:The present invention provides a new approach to creating novel polynucleotide sequences by point mutation and recombination in vitro of a set of parental sequences. The new polynucleotide sequences can be useful in themselves or they can be used for the templated synthesis of polymers, e.g. polypeptides composed of α-amino acids as occurring naturally in protein synthesis or polymers comprised of β-amino acids or other building blocks as described (Templated molecules).
专利号:US-2007059711-A1 优先权日:2004-07-30 标 题:Microarray for evaluation of stress-related genes in skin 发明人:SLOMINSKI ANDRZEJ; PISARCHIK ALEXANDER 权利人:SLOMINSKI ANDRZEJ; PISARCHIK ALEXANDER 摘要:The present invention provides a novel DNA microarray chip that can be used for simultaneous testing of transcriptional responses to cutaneous stressors in the context of neuro-endocrine-immune functions of the skin. The transcriptional responses to ultraviolet radiation in epidermal keratinocytes were tested using such microarray chip containing more than 700 neuro-endocrine-immune related genes. The gene expression pattern was non-random and time dependent; it included increased expression of genes involved in water and salt balance, prostaglandin synthesis, keratinocyte differentiation as well as genes coding for stress effectors, cytokines and metalloproteinases. In contrast, expression was decreased for genes coding for growth factors and their receptors, and for elements of extracellular matrix. This stochastic pattern suggests that transcriptional responses are coordinated and aimed at preservation of epidermal barrier function, prevention of early carcinogenic events and remodeling of extracellular matrix.
参考文献:10.1016/s0076-6879(05)07013-8 摘要:Svensson AW, Casey PJ, Young SG, Bergo MO. Genetic and pharmacologic analyses of the role of Icmt in Ras membrane association and function. Methods Enzymol. 2006;407():144–59. doi: 10.1016/s0076-6879(05)07013-8.