144 results about "Cloning genes" patented technology

The production of a diutan polysaccharide exhibiting increased viscosity properties as compared with previously produced polysaccharide of the same type of repeating units. Such an improved diutan polysaccharide is produced through the generation of a derivative of Sphingomonas sp. ATCC 53159 that harbors a multicopy broad-host-range plasmid into which genes for biosynthesis of diutan polysaccharide have been cloned. The plasmid provides the capability within the host Sphingomonas strain to produce multiple copies of genes for such polysaccharide synthesis. In such a manner, a method of not just increased production of the target diutan polysaccharide, but also production of a diutan polysaccharide of improved physical properties (of the aforementioned higher viscosity) thereof is provided. Such a diutan polysaccharide has proven particularly useful as a possible viscosifier in oilfield applications and within cement materials. The inventive methods of production of such an improved diutan polysaccharide, as well as the novel cloned genes required to produce the improved diutan within such a method, are also encompassed within this invention. Additionally, the novel engineered Sphingomonas strain including the needed DNA sequence is encompassed within this invention.

The present invention is a substantially purified sortase-transamidase enzyme from Gram-positive bacteria, such as Staphylococcus aureus. The enzyme having a molecular weight of about 23,539 or about 29,076 daltons and catalyzing a reaction that covalently cross-links the carboxyl terminus of a protein having a sorting signal to the peptidoglycan of a Gram-positive bacterium, the sorting signal having: (1) a motif of LPX3X4G therein; (2) a substantially hydrophobic domain of at least 31 amino acids carboxyl to the motif; and (3) a charged tail region with at least two positively charged residues carboxyl to the substantially hydrophobic domain, at least one of the two positively charged residues being arginine, the two positively charged residues being located at residues 31-33 from the motif, wherein X3 is any of the twenty naturally-occurring L-amino acids and X4 is selected from the group consisting of alanine, serine, and threonine, and wherein sorting occurs by cleavage between the fourth and fifth residues of the LPX3X4G motif. Variants of the enzyme, methods for cloning the gene encoding the enzyme and expressing the cloned gene, and methods of use of the enzyme, including for screening for antibiotics and for display of proteins or peptides on the surfaces of Gram-positive bacteria, are also disclosed.

The invention discloses a salvia miltiorrhiza bunge isopentenylpyrophosphate isomerase (IPPI) gene, a protease coded by the IPPI genes and an application thereof. The gene is obtained by cloning fromsalvia miltiorrhiza bunge through utilizing a cDNA chip technology for the first time, thus the invention fills the blank of separating and cloning the SmIPPI gene from the salvia miltiorrhiza bunge which is a conventional famous and precious Chinese medicinal plant. The SmIPPI gene has a nucleotide sequence represented by SEQ ID NO.1 or adds, substitutes, inserts or deletes one or more nucleotidehomologous sequences or an allele thereof and a nucleotide sequence derived from the nucleotide homologous sequence. A protein coded by the SmIPPI genes has an amino acid sequence represented by SEQID NO.2 or adds, substitutes, inserts or deletes one or more amino acid homologous sequences. The SmIPPI gene can be applied to researches and industrialization for improving the content of diterpeneactive constituents in the salvia miltiorrhiza bunge by a biotechnology method and improving the content of tanshinone substances by utilizing a transgenic technology, is helpful to the quality improvement, the selective breeding, and the like of medicinal materials of the salvia miltiorrhiza bunge and has good application prospect.

The invention discloses application of SBP-box type transcription factors of switchgrass in the aspect of increasing plant biomass and fermentable sugar yields, and belongs to the technical field of plant genetic engineering. The application mainly includes cloning genes of the SBP-box type transcription factors PvSPL1 and 2 of the switchgrass and determining sequences of the SBP-box type transcription factors of the switchgrass; inhibiting the transcription activity of integral subfamilies of the transcription factors PvSPL1 and 2 on the basis of the genes and the sequences by the aid of chimeric repressor silencing technologies (CRES-T) so as to generate transgenic switchgrass plants with increased tiller quantities. The application has the advantages that dry matter biomass of the obtained transgenic switchgrass plants can be increased by 2.0-2.3 times, and the total fermentable sugar yields can be increased by 2.0-2.2 times; the generated transgenic switchgrass energy plants can be integrated in conventional breeding projects, and accordingly precious germplasm resources can be provided for creating novel varieties of high-yield and efficiently degradable energy plants; novel targets for molecular breeding in future can be provided by the authenticated SBP-box type transcription factors PvSPL1 and 2 of the switchgrass, and the SBP-box type transcription factors PvSPL1 and 2 of the switchgrass can be used for designing oriented molecules of commercialized energy plant products.

The present invention relates to the construction and utilization of a new mammalian expression vector that contains a unique multiple cloning site (MCS), designated pUHAB. The pUHAB vector comprises a high copy replication origin (ColE1), a drug resistance gene (TK-Hygromycin), and a human cytomegalovirus promoter operably associated with a unique intron (hCMV / intron). Further, pUHAB comprises a selectable marker conferring resistance to kanamycin in bacterial cells, and a phage f1(+) region. pUHAB can be used to transiently or stably express cloned genes when transfected into mammalian cells. The invention also encompasses kits and host cells and cell lines comprising pUHAB, and methods of producing a recombinant protein using pUHAB.

The invention belongs to the technical field of the livestock genetic engineering and in particular is a molecular marker which is used as the application to pig marker-assisted selection and relative with immune traits and the application. The molecular marker is obtained by cloning gene BCL10, the nucleotide sequence of the molecular marker is displayed as the sequence table SEQ ID NO: 2, and the number 144 basic group position of the sequence which is displayed in the sequence table SEQ ID NO: 2 has a gene mutation of C144-T144, which leads to the polymorphism of the RFLP-HinP1I. The invention also discloses a process for preparing the molecular marker and the application of a polymorphism testing method and provides a new molecular marker for the pig marker-assisted selection.

The fast cucurbitaceae RIP gene screening method includes designing degenerate primer pair LY1 / LY2 based on highly conservative amino acid sequence and its codon bias in the upstream and downstream of known cucurbitaceae RIP, Touchdown-PCR amplification with total DNA as template, direct PCR product clonal sequencing of cucurbitaceae plant with un-reported RIP gene to obtain new RIP gene; and fast distinguishing known and unknown RIP genes of cucurbitaceae plant with reported RIP gene according to the length polymorphism of limiting segment in RIP gene LY1 / LY2 segments. Compared with available method of first separating RIP and then cloning gene, the method of the present invention has the features of being fast and accurate, and may be used widely in RIP gene screening.

Glycerol-3-phosphate acyltransferase is the initial enzyme of the glycerolipid biosynthetic pathway. Biochemical analyses indicated that the reaction mediated by glycerol-3-phosphate acyltransferase represents a potential rate-limiting step for the synthesis of phospholipids and storage neutralipid, triacylglycerol. The present invention relates to the cloning of genes encoding extraplastidic membrane-bound glycerol-3-phosphate acyltransferases. Heterologous expression of the genes, GPAT1, GPAT2, and GPAT3 in a yeast glycerol-3-phosphate acyltransferase mutant demonstrated that the encoded products could efficiently utilize glycerol-3-phosphate to mediate sn-1 stereo-specific fatty acid acylation. The invention encompasses the glycerol-3-phosphate acyltransferase peptides disclosed and fragments and homologues thereof, the corresponding gene sequences and fragments and homologues thereof, as well as the use of the peptide and gene sequences of the present invention for use in generating recombinant proteins, and transgenic plants with altered lipid metabolism. In this way, the present invention also encompasses the use of such recombinant peptides and transgenic plants for the production of lipid products for use, for example, in pharmaceutical and nutritional applications.

The objective of the invention is to provide a method for applying miRNAs or miRNA clusters in changing cell fates by promoting the formation of induced pluripotent stem cells. The method comprises a first step of cloning genes of miRNAs or miRNA clusters in a donor genome and obtaining a donorcell, a second step of changing the cell fate of the donorcell in step 1 by using the miRNAs or miRNA clusters in step 1, and a third step of identifying the cell obtained in step 2. Compared to the prior art, stem cells are induced by miRNAs or miRNA clusters in the invention instead of by oncogenes c-Myc, thereby reducing potential carcinogenicity, accelerating the change of cell fates and enabling a high induction efficiency and high-quality induced pluripotent stem cells with ability of germ line transmission to be obtained.