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Showing 1 to 18 of 18 for “"subcloning"”.
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Subcloning and regulation of a human intestinal sodium-phosphate cotransporter gene
Phosphate plays a critical role in the body as a constituent of bone and tooth for body development, and as a urinary buffer for pH in body acid-base balance regulation. The phosphorus level in blood in human is between 3.0 to 4.5 mg/dl. When the blood phosphate concentration is lower than 2.5 …
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Genetic engineering of recombinant anti-mycolic acid antibody fragments for use in tuberculosis diagnostics
… were investigated as possible targets for subcloning. The plasmids pGE20 and pAK400 were previously used for the expression of scFv antibody proteins. Subcloning into plasmid pAK400 proved to be the more efficient of the two investigated for subcloning. This subcloning yielded the …
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Genetic and Biochemical Characterization of the Lipid-Protein Interactions of Pyruvate Oxidase
… no direct selection was available. Subsequent subcloning resulted in localization of the gene to a 3.2-kilobase-pair DNA segment, which was shown to cause the overproduction of oxidase activity (by six- to eightfold) and encode a protein having the size and antigenic determinants of pyruvate …
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Chemical synthesis, cloning and expression of a gene encoding systemin, a proteinase inhibitor-inducing factor
… was cloned into pUCJ 8 for amplification and subcloning into pMAL-pk for expression as a maltose binding-fusion protein. The recombinant systemin was released by enterokinase and isolated by HPLC. After further purification, the physical characteristics including amino acid composition, …
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Transfektion migrierender Nierenepithelzellen mit dem Kalium-Kanal ROMK2
… Transfizierte Zellklone konnten durch Subcloning und RT-PCR-Experimente mit spezifischen ROMK2-Primern gefun- den werden. Desweiteren wurden Patch-Clamp-Experimente im Ganzzellmodus durch- geführt, um die Insertion von funktionellen ROMK2-Kanälen in die Zellmembran von transfizierten …
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Cloning, sequencing and physicochemical characterization of a hemagglutinin from Escherichia coli 09:H10:K99
… was localized and identified unambiguously by subcloning. The nucleotide sequence of the gene was determined and found to consist of a 792 base pair open reading frame coding for a protein of 29 kDa. This protein has a predicted prokaryotic N-terminal secretory signal sequence. The protein …
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Identification and Characterization of the Cis-acting Elements Around the Murine cd4 Cnhancer
… silencer functions were further confirmed by subcloning them at different positions and different orientations relative to CD4 promoter. These subcloned constructs were tested for function in transient transfection and CAT assays. Furthermore these two <em>cis</em>-acting elements were also …
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Identification and characterisation of microRNAs expressed in articular cartilage and osteoarthritis
… from the array were further validated by subcloning and luciferase assays, resulting in the discovery of ITGA5 as a target of candidate novel 11. The outcomes from this research provide us with an increased understanding of the miRNome of human OA cartilage and could impact upon future …
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Gene cloning, adjustable expression and site-directed mutagenesis of Escherichia coli adenylate kinase
… been cloned by phage Mu-mediated transposition. Subcloning of the adk$\sp{+}$ gene into pUC13 and pBR322 allowed high-level expression and a simple purification scheme for the enzyme. An optimized system was developed for the in vivo transfer of a mutant gene from the Escherichia coli chromosome …
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Characterization of cytoplasmic diversity in soybean (Glycine max L. Merr) using mitochondrial markers
… also detected. The second objective included subcloning and sequencing the 0.9 kb and 1.7 kb <i>Hind</i>III-<i>Pst</i>I clones flanking the 2.3 kb <i>Hind</i>III fragment from 'Williams 82' DNA The total 4.9 kb <i>Pst</i>I sequence from 'Williams 82' mitochondrial DNA was used to search a …
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Characterization of a human renal organic anion transporter
… an hROAT1 expression clone was constructed by subcloning the hROAT1 reading frame between the 5' and 3' UTRs of the flounder sodium dicarboxylate cotransporter, fNaDC-3. Expressed in Xenopus laevis oocytes, hROAT1 mediated the uptake of p-aminohippurate (PAH), the model substrate for this …
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Development and characterization of recombinant human autoantibody using cDNA made from peripheral blood mononuclear cells of patients with bronchial asthma
… ELISA를 통한 CK-18 단백질 특이 scFv의 선별 8 3. scFv정제를 위한 subcloning 9 4. scFv 단백질 정제 10 E. 선별된 scFv항체의 CK-18 단백질에 대한 반응 평가 11 1. Sodiumdodecylsµlfate-polyacrylamide gel electrophoresis (SDS-PAGE) 와 immunoblot 11 2. 정제된 scFv항체들을 이용한 ELISA 11 F. 인간 유전자 재조합 단클론 IgG 자가항체의 제작 및 정제 12 1. IgG 자가항체의 제작 12 …
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Evolution of a gene for pathogenicity: endo-pectate lyase
… endo-pectate lyase gene from Ecc was chosen for subcloning and further study. I used the plasmid vector pBR322 to construct a clone bank of Eca strain SRB; of the 1700 clones screened, five were pectolytic. Two of the Eca pectolytic. clones had homology to the Ecc endo-pectate lyase gene; upon …
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Mechanisms of dendritic peptide release
… under different experimental conditions. Subcloning of a red fluorescent construct tagged to ppANF was tested on PC12 cells to show targeting of fluorescence expression to LDCVs. Successful production of an inducible adenoviral DNA with the red fluorescent construct insert was confirmed by …
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Natural Products From Functional Screening of Soil Metagenomic Libraries
… and confirmed using transposon mutagenesis and subcloning. These antibacterial activities could not be replicated when clones were transformed into Escherichia coli, indicating the powerful ability of R. metallidurans to find diverse antibacterially active enzymes in metagenomic screens. …
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MODULATION OF THE B-CELL REPERTOIRE IN RHEUMATOID ARTHRITIS BY TRANSIENT B-CELL DEPLETION
… using family-specific primers, followed by subcloning and sequencing. During the study, patient H received two courses of antibody treatment. B-cell depletion lasted 7 and 10 months, respectively and each time was accompanied by a clinical improvement. Anti-CD20 therapy induced two types of …
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ΜΕΛΕΤΗ ΤΩΝ ΦΥΣΙΚΩΝ ΠΛΑΣΜΙΔΙΩΝ ΤΟΥ ΣΤΕΛΕΧΟΥΣ ATCC 10988 ΤΟΥ ΑΙΘΑΝΟΛΟΠΑΡΑΓΩΓΟΥ ΒΑΚΤΗΡΙΟΥ ZYMOMONAS MOBILIS
FIVE NATIVE PLASMIDS OF Z. MOBILIS STRAIN ATCC 10988 HAVE BEEN ISOLATED, NAMELYPZMO2, PZMO3, PZMO5, PZMO6 WITH SIZE 1,70, 1,85, 2,70, 16,7, AND 31,8 KB RESPECTIVELY. RESTRICTION MAPPING (PARTIAL) OF ALL THESE PLASMIDS WAS FOLLOWED AND THEIR HOMOLOGY LEVELS WERE DETERMINED. SIMILARLY, THEIR HOMOLOGY …