Chapter 20
... laboratory share general features, such as the use of bacteria and their plasmids • Plasmids are small circular DNA molecules that replicate separately from the bacterial chromosome ...
... laboratory share general features, such as the use of bacteria and their plasmids • Plasmids are small circular DNA molecules that replicate separately from the bacterial chromosome ...
Genetic data indicate that proteins containing the GGDEF domain
... pYhcK £occulated heavily (Fig. 3B,C). Strong £uorescence was observed in and around these aggregates in the microscope after staining with Calco£uor (data not shown). Plasmid pYhcK caused the strongest aggregation in the liquid culture (Fig. 3C). These results support the data presented by Amikam an ...
... pYhcK £occulated heavily (Fig. 3B,C). Strong £uorescence was observed in and around these aggregates in the microscope after staining with Calco£uor (data not shown). Plasmid pYhcK caused the strongest aggregation in the liquid culture (Fig. 3C). These results support the data presented by Amikam an ...
Document
... identical copies of a gene-carrying piece of DNA – Recombinant DNA is formed by joining DNA sequences from two different sources – One source contains the gene that will be cloned – Another source is a gene carrier, called a vector ...
... identical copies of a gene-carrying piece of DNA – Recombinant DNA is formed by joining DNA sequences from two different sources – One source contains the gene that will be cloned – Another source is a gene carrier, called a vector ...
Protein expression, purification, and molecular cloning
... When adequate separation of bands has occurred, carefully excise the DNA fragment of interest using a wide, clean, sharp scalpel. Minimize the size of the gel slice by removing extra agarose. Determine the appropriate volume of the gel slice by weighing it in a clean 1.5 mL microcentrifuge tube. Ass ...
... When adequate separation of bands has occurred, carefully excise the DNA fragment of interest using a wide, clean, sharp scalpel. Minimize the size of the gel slice by removing extra agarose. Determine the appropriate volume of the gel slice by weighing it in a clean 1.5 mL microcentrifuge tube. Ass ...
Restriction enzymes
... • These enzymes protect bacteria against intruding DNA from other organisms. • They work by cutting up the foreign DNA, a process called restriction. • If foreign DNA enters the bacteria cell the restriction enzyme will cut it up into small pieces. They cut up only certain base pair sequences and th ...
... • These enzymes protect bacteria against intruding DNA from other organisms. • They work by cutting up the foreign DNA, a process called restriction. • If foreign DNA enters the bacteria cell the restriction enzyme will cut it up into small pieces. They cut up only certain base pair sequences and th ...
Methods S1: Vector constructions and transformation of yeast and
... AvrLm4-7 in tobacco leaves, alone or fused to eGFP. eGFP coding sequence was amplified from plasmid peGFP (Clontech, Mountain View, CA, USA) using primers pBINeGFPXbaUp (which introduces a XbaI restriction site) and pBINeGFP-SacLo (which introduces a SacI restriction site). eGFP PCR product was dige ...
... AvrLm4-7 in tobacco leaves, alone or fused to eGFP. eGFP coding sequence was amplified from plasmid peGFP (Clontech, Mountain View, CA, USA) using primers pBINeGFPXbaUp (which introduces a XbaI restriction site) and pBINeGFP-SacLo (which introduces a SacI restriction site). eGFP PCR product was dige ...
Presentazione di PowerPoint
... • It is updated daily and a coordination with NCBI and DDBJ ensures its completeness • It is offered at EBI by means of SRS ...
... • It is updated daily and a coordination with NCBI and DDBJ ensures its completeness • It is offered at EBI by means of SRS ...
Enzyme Mechanisms - Illinois Institute of Technology
... Gel soaked in base to denature duplexes pH readjusted to neutral Sheet of absorbent material placed atop the gel Salt solution is drawn across the gel, perp to the electrophoretic direction, in various ways to carry the DNA onto the sheet Sheet is dried in an oven to tightly attach the DNA to it Inc ...
... Gel soaked in base to denature duplexes pH readjusted to neutral Sheet of absorbent material placed atop the gel Salt solution is drawn across the gel, perp to the electrophoretic direction, in various ways to carry the DNA onto the sheet Sheet is dried in an oven to tightly attach the DNA to it Inc ...
video slide
... Each sample, a mixture of DNA molecules, is placed in a separate well near one end of a thin slab of gel. The gel is supported by glass plates, bathed in an aqueous solution, and has electrodes attached to each end. When the current is turned on, the negatively charged DNA molecules move toward the ...
... Each sample, a mixture of DNA molecules, is placed in a separate well near one end of a thin slab of gel. The gel is supported by glass plates, bathed in an aqueous solution, and has electrodes attached to each end. When the current is turned on, the negatively charged DNA molecules move toward the ...
Slide 1 - Schools
... identical copies of a gene-carrying piece of DNA – Recombinant DNA is formed by joining DNA sequences from two different sources – One source contains the gene that will be cloned – Another source is a gene carrier, called a vector ...
... identical copies of a gene-carrying piece of DNA – Recombinant DNA is formed by joining DNA sequences from two different sources – One source contains the gene that will be cloned – Another source is a gene carrier, called a vector ...
Biosynthesis of the Antibiotic Nonribosomal Peptide Penicillin in
... Many important therapeutics including key antibiotics are derived from compunds produced by fungal organisms1, yet fungal enzymatic diversity is a largely untapped resource for cheap biosynthesis of medical molecules and the potential for the discovery of novel therapeutics2, 3. The baker’s yeast Sa ...
... Many important therapeutics including key antibiotics are derived from compunds produced by fungal organisms1, yet fungal enzymatic diversity is a largely untapped resource for cheap biosynthesis of medical molecules and the potential for the discovery of novel therapeutics2, 3. The baker’s yeast Sa ...
Recombinant Technology
... 12.5 Reverse transcriptase helps make genes for cloning • Reverse transcriptase can be used to make smaller, complementary DNA (cDNA) libraries ...
... 12.5 Reverse transcriptase helps make genes for cloning • Reverse transcriptase can be used to make smaller, complementary DNA (cDNA) libraries ...
Genetic Basis of Variation in Bacteria Genetic Basis of Variation in
... Organization of genetic material in bacteria: plasmids Examples of naturally-occuring plasmids and relevant features ...
... Organization of genetic material in bacteria: plasmids Examples of naturally-occuring plasmids and relevant features ...
Jeopardy
... Question: Which of the following is an example of successful transformation? a. Injection of bacterial DNA into plant cells. b. A defective gene in a cell being replaced with a normal gene c. A bacterial cell that expresses the lactase gene in the presence of lactose d. None of the above BACK TO GAM ...
... Question: Which of the following is an example of successful transformation? a. Injection of bacterial DNA into plant cells. b. A defective gene in a cell being replaced with a normal gene c. A bacterial cell that expresses the lactase gene in the presence of lactose d. None of the above BACK TO GAM ...
Isolation and Characterization of Mutations in the b-Tubulin Gene of Saccharomyces cerevisiae .
... for 60 min at 30" with gentle rocking. The resulting spheroplasts were centrifuged for 10 sec in a microfuge, and the pellet was resuspended gently in 0.5 ml of 50 mM EDTA, 0.3% SDS, pH 8.5. T h e tube was heated to 65" for 20 min, then 100 pI of 5 M potassium acetate was added, chilled on ice for 2 ...
... for 60 min at 30" with gentle rocking. The resulting spheroplasts were centrifuged for 10 sec in a microfuge, and the pellet was resuspended gently in 0.5 ml of 50 mM EDTA, 0.3% SDS, pH 8.5. T h e tube was heated to 65" for 20 min, then 100 pI of 5 M potassium acetate was added, chilled on ice for 2 ...
The pyruvate dehydrogenase complex of the chemol
... which lacks pyruvate decarboxylase, dihydrolipoyl transacetylase and dihydrolipoamide dehydrogenase activities and is unable to grow on minimal medium lacking acetate whereas PDH positive strains can (Guest et al., 1983). Cosmid 5.1, plasmids pTHI081 and pTHI0251 were transformed into E. coli JRG134 ...
... which lacks pyruvate decarboxylase, dihydrolipoyl transacetylase and dihydrolipoamide dehydrogenase activities and is unable to grow on minimal medium lacking acetate whereas PDH positive strains can (Guest et al., 1983). Cosmid 5.1, plasmids pTHI081 and pTHI0251 were transformed into E. coli JRG134 ...
PART I
... pairing of a few bases, known as microhomologies, are required for a pre-annealing step between T-DNA strand coupled with VirD2 and plant DNA. The microhomologies can provide a minimum specificity required for the recombination process. The Ti system probably inserts only one T-DNA segment per bacte ...
... pairing of a few bases, known as microhomologies, are required for a pre-annealing step between T-DNA strand coupled with VirD2 and plant DNA. The microhomologies can provide a minimum specificity required for the recombination process. The Ti system probably inserts only one T-DNA segment per bacte ...
PRINCIPLES OF RECOMBINANT DNA TECHNOLOGY
... Essentially there are three types of vectors used in this work: plasmids, phages and cosmids. All replicate within the host bacterial cell and are therefore sometimes referred to as replicons. Plasmids occur naturally in bacteria to which they confer resistance to various antibiotics, heavy metals, ...
... Essentially there are three types of vectors used in this work: plasmids, phages and cosmids. All replicate within the host bacterial cell and are therefore sometimes referred to as replicons. Plasmids occur naturally in bacteria to which they confer resistance to various antibiotics, heavy metals, ...
Molecular Plant-Microbio Interactions
... flanking regions were sequenced and used to perform searches in protein and nonredundant nucleotide databases. No significant similarity or homology with any known sequence was detected. Additional keywords: host specificity, N2 fixation, R. leguminosarum bv. viciae, transposon. Formation of nitroge ...
... flanking regions were sequenced and used to perform searches in protein and nonredundant nucleotide databases. No significant similarity or homology with any known sequence was detected. Additional keywords: host specificity, N2 fixation, R. leguminosarum bv. viciae, transposon. Formation of nitroge ...
Gene cloning of P43 surface protein of toxoplasma gondii tachyzoite
... depends on the stage of the pregnancy period which the infection occurs (Zhao 1992; Wallon et al, 2002). This parasite will be detected in human beings by serological tests only, and specific antigen is very essential in diagnosis system. P43 (SAG3) is one member of the redundant system of T. gondii ...
... depends on the stage of the pregnancy period which the infection occurs (Zhao 1992; Wallon et al, 2002). This parasite will be detected in human beings by serological tests only, and specific antigen is very essential in diagnosis system. P43 (SAG3) is one member of the redundant system of T. gondii ...
A Tn 10-lacZ-kanR-URA3 Gene Fusion Transposon for Insertion Mutagenesis and Fusion Analysis of Yeast and Bacterial Genes.
... and extracting plasmid DNA (with o r without a short period of growth). In such a DNA preparation, approximately 2% of the target plasmid molecules contain a transposon insertion, as judged by transformation and selection for AmpR followed by screening for kanamycin resistance; in our analysis of LE ...
... and extracting plasmid DNA (with o r without a short period of growth). In such a DNA preparation, approximately 2% of the target plasmid molecules contain a transposon insertion, as judged by transformation and selection for AmpR followed by screening for kanamycin resistance; in our analysis of LE ...
NucleoSpin 96 Flash Plasmid and Large-Construct DNA
... Note: Direct inoculation of cultures for BAC DNA preparation from single colonies or glycerol stocks (without preculture) may result in lower yields and less reproducible results due to higher differences in yields. Use a suitable pin-tool for 96-well plates or 8-channel pipette to inoculate the cul ...
... Note: Direct inoculation of cultures for BAC DNA preparation from single colonies or glycerol stocks (without preculture) may result in lower yields and less reproducible results due to higher differences in yields. Use a suitable pin-tool for 96-well plates or 8-channel pipette to inoculate the cul ...
PCR-based gene synthesis to produce recombinant proteins for
... Design of oligonucleotides for gene synthesis Oligonucleotides for gene synthesis were designed using DNAWorks [3,4] with the following parameters: E. coli class II codon frequency table, 60°C annealing temperature, 60 (for polA) and 75 (for paz) nt oligonucleotide length, 5 solutions and TBIO mode. ...
... Design of oligonucleotides for gene synthesis Oligonucleotides for gene synthesis were designed using DNAWorks [3,4] with the following parameters: E. coli class II codon frequency table, 60°C annealing temperature, 60 (for polA) and 75 (for paz) nt oligonucleotide length, 5 solutions and TBIO mode. ...
Transformation Lab - Towson University
... encoded in the foreign DNA. Transformation enables inexpensive and reliable production of important medical products such as insulin, human growth hormone, and other replacement hormone and gene therapies. In bacteria, the haploid genome is a single circular chromosome. This differs from eukaryotic ...
... encoded in the foreign DNA. Transformation enables inexpensive and reliable production of important medical products such as insulin, human growth hormone, and other replacement hormone and gene therapies. In bacteria, the haploid genome is a single circular chromosome. This differs from eukaryotic ...
Plasmid
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A plasmid is a small DNA molecule within a cell that is physically separated from a chromosomal DNA and can replicate independently. They are most commonly found in bacteria as small, circular, double-stranded DNA molecules; however, plasmids are sometimes present in archaea and eukaryotic organisms. In nature, plasmids often carry genes that may benefit the survival of the organism, for example antibiotic resistance. While the chromosomes are big and contain all the essential information for living, plasmids usually are very small and contain only additional information. Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms.Plasmids are considered replicons, a unit of DNA capable of replicating autonomously within a suitable host. However, plasmids, like viruses, are not generally classified as life. Plasmids can be transmitted from one bacterium to another (even of another species) via three main mechanisms: transformation, transduction, and conjugation. This host-to-host transfer of genetic material is called horizontal gene transfer, and plasmids can be considered part of the mobilome. Unlike viruses (which encase their genetic material in a protective protein coat called a capsid), plasmids are ""naked"" DNA and do not encode genes necessary to encase the genetic material for transfer to a new host. However, some classes of plasmids encode the conjugative ""sex"" pilus necessary for their own transfer. The size of the plasmid varies from 1 to over 200 kbp, and the number of identical plasmids in a single cell can range anywhere from one to thousands under some circumstances.The relationship between microbes and plasmid DNA is neither parasitic nor mutualistic, because each implies the presence of an independent species living in a detrimental or commensal state with the host organism. Rather, plasmids provide a mechanism for horizontal gene transfer within a population of microbes and typically provide a selective advantage under a given environmental state. Plasmids may carry genes that provide resistance to naturally occurring antibiotics in a competitive environmental niche, or the proteins produced may act as toxins under similar circumstances, or allow the organism to utilize particular organic compounds that would be advantageous when nutrients are scarce.