Plasmid construction, cloning, and cell strains
Cloning Nanobody cDNA into pHEN6c Plasmid Vector
... Only transformed bacteria that contain the plasmid and express β-lactamase grew on plates which contain ampicillin [10]. Untransformed cells like the plate number two of our sample could not grow on the ampicillin ...
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Rapid Construction of Recombinant Plasmids by QuickStep-Cloning
... molecular cloning technique that builds upon the concepts of asymmetric PCR and megaprimer-based amplification of ...a plasmid of choice, in a time-efficient and cost-effective manner, without the need of a ...
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Cloning of a Recombinant Plasmid Encoding Thiol-Specific Antioxidant Antigen (TSA) Gene of Leishmania major and Expression in the Chinese Hamster Ovary Cell Line
... pcDNA3 plasmid, the gene was cloned with linkers to join it to the HindIII and EcoRI sites of pcDNA3 (Invitrogen, US) to produce the recombinant eukaryotic expression plasmid ...The plasmid was then ...
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Generation of Mutant Murine Cytomegalovirus Strains from Overlapping Cosmid and Plasmid Clones
... overlapping plasmid subclones. A new vector, PmeSUB, featuring a multiple cloning site and a stringent origin of replication, was constructed to make large subclones for use with smaller subclones ...
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Cloning, expression, purification and characterization of replication protein from plasmid pGP2 from Acetobacter estunensis
... 1.3. Cloning and Expression Vector Construction Recombinant DNA techniques were performed using conventional protocols. The rep 34 gene of Acetobacter estunensis (ORF2) was amplified using: forward 5´-GGA ...
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Cloning and Expression of Recombinant Plasmid Containing P36/LACK Gene of Leishmania infantum Iranian Strain.
... The extracted DNA was used as a template to amplify the LACK gene by PCR. The reac- tion was performed in 100μl of the solution containing: 1μl of template DNA, 2μl of dNTP (100 μmol), 0.5μl of Taq DNA poly- merase ...
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Construction and Characterization of a Replication-Competent Retroviral Shuttle Vector Plasmid
... We particularly wanted to develop a vector system that makes it simple to recover both unintegrated and integrated DNA. To simplify enrichment of viral DNA, we included the lacO sequence in the inserted segment. DNAs ...
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Construction of Mtb72F Plasmid as a DNA Vaccine Candidate for Mycobacterium tuberculosis
... Mtb72F consists of the C- and N-terminal open reading frames (ORFs) of Mtb32 (Rv0125) with the mtb39 gene (Rv1196) between them. The resulting recombinant vaccine, Mtb72F, provided greater protection against TB in ...
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Characterization of Shigella Strains in Iran by Plasmid Profile Analysis and PCR Amplification of ipa Genes
... during cell storage and subculturing or plasmid ...of plasmid DNA revealed that all 82 isolates harbored multiple plasmids, with an average of ...all strains and a mean of 10, 8, 10, and 9 ...
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Molecular cloning of the Aleutian disease virus genome: expression of Aleutian disease virus antigens by a recombinant plasmid.
... by nick translation, DNA relatedness to several field strains of ADV (Utah I, Pullman, and DK), but not to mink enteritis virus or cellular DNA, was shown by Southern blot hybridization.[r] ...
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For the construction of the Egr1 expression plasmid, the pcmv-sport6-egr1 vector (Invitrogen),
... Deparaffinization, endogenous peroxidase inactivation, antigen retrieval from formalin fixed paraffin embedded clinical tissues, and immunostaining of Brm was performed as described previously (14). For Egr1 ...
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Construction of pRSET-sfGFP Plasmid for Fusion-Protein Expression, Purification and Detection
... the construction of the protein expression plasmid pRSET-sfGFP by cloning sfGFP gene downstream of the N-terminal 6×His tag in the T7 promoter-plasmid ...this plasmid in the cytoplasm ...
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Molecular Cloning, Expression and Characterization of Plasmid Encoding Rhomboid 4 (ROM4) of Tachyzoite of Toxoplasma gondii RH Strain
... There is no effective vaccine in humans and Toxovax (S48 strain) is the only accessible commercial vaccine in sheep licensed in Eu- rope and New Zealand but there is safety con- cern due to recrudescence of the infection ...
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Construction of a self-cloning system in the unicellular green alga Pseudochoricystis ellipsoidea
... Yuki Kasai 1* , Kohei Oshima 1 , Fukiko Ikeda 1 , Jun Abe 1 , Yuya Yoshimitsu 1,2 and Shigeaki Harayama 1 Abstract Background: Microalgae have received considerable interest as a source of biofuel production. The ...
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Rapid construction of mycobacterial mutagenesis vectors using ligation independent cloning
... The construction of targeted mycobac- terial mutants has been invaluable for the understanding of gene function, the demonstration of gene essentiality and in the generation of attenuated strains for ...
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Designing and Construction of a Cloning Vector Containing mpt64 Gene of Mycobacterium tuberculosis
... pcDNA3.1 (+) vector, 2.5 μL T4 DNA ligase buffer, 0.2 μL T4 DNA ligase (150 U/μL) (Metabion, Germany) and 2 μL of the polyethylene glycol (PEG) 4000 (Thermo Scientific, Germany). After ligation, the competent cells of ...
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Design and Construction of a Cloning Vector Containing the hspX Gene of Mycobacterium tuberculosis
... Background: Tuberculosis (TB) is a major cause of death worldwide. Finding an effective vaccine against TB is the best way to control it. Several vaccines against this disease have been developed but none are completely ...
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Utilising the native plasmid, pCA2.4, from the cyanobacterium Synechocystis sp. strain PCC6803 as a cloning site for enhanced product production
... native plasmid when compared to the isogenic chromosomally inte- grated control ...small plasmid, ...PCC6803 strains having high level, controlled expression may be more ...
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Construction of Escherichia coli Strains for Conversion of Nitroacetophenones to ortho-aminophenols
... constructed strains might accumulate the aminophenols from nitroacetophenones and other nitroaromatic ...recombinant plasmid carrying NB nitroreductase (nbzA) and HAB mutase A (habA) genes, both from ...
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Construction of Bacillus thuringiensis simulant strains suitable for environmental release
... insertion plasmid and grown on LB agar plates containing spectinomycin at ...expression plasmid pAD02, and then grown on LB agar plates containing tetracycline at ...
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