Isolation of insertion elements from Gram
... in question into E. coli and by testing for sucrose sensitivity (data not shown). Therefore, besides mutation of the sacB gene, at least one other, so far unknown resistance mechanism should exist. Since in E. coli the lethal effect parallels with an active levan sucrase in the periplasm [2], chromo ...
... in question into E. coli and by testing for sucrose sensitivity (data not shown). Therefore, besides mutation of the sacB gene, at least one other, so far unknown resistance mechanism should exist. Since in E. coli the lethal effect parallels with an active levan sucrase in the periplasm [2], chromo ...
GROW`N`GLOW: THE ACE1 TWO-HYBRID
... valuable addition to the family of reporter genes for two-hybrid systems and makes a secondary screening of yeast colonies faster and more cost-effective than conventional LacZ assays. Detection of protein-protein interactions via the green fluorescent protein provides global screening of colonies w ...
... valuable addition to the family of reporter genes for two-hybrid systems and makes a secondary screening of yeast colonies faster and more cost-effective than conventional LacZ assays. Detection of protein-protein interactions via the green fluorescent protein provides global screening of colonies w ...
Structure and Physiological significance of lipid
... 4. Introduction of recombinant DNA into compatible host cells. • In order to be propagated, the recombinant DNA molecule (insert DNA joined to vector DNA) must be introduced into a compatible host cell where it can replicate. • The direct uptake of foreign DNA by a host cell is called genetic trans ...
... 4. Introduction of recombinant DNA into compatible host cells. • In order to be propagated, the recombinant DNA molecule (insert DNA joined to vector DNA) must be introduced into a compatible host cell where it can replicate. • The direct uptake of foreign DNA by a host cell is called genetic trans ...
Recombinant DNA Technology
... Identification of host cells containing recombinant DNA requires genetic selection or screening or both. In a selection, cells are grown under conditions in which only transformed cells can survive; all the other cells die. In contrast, in a screen, transformed cells have to be individually tested f ...
... Identification of host cells containing recombinant DNA requires genetic selection or screening or both. In a selection, cells are grown under conditions in which only transformed cells can survive; all the other cells die. In contrast, in a screen, transformed cells have to be individually tested f ...
Lecture NoteIV
... plasmid DNA during the plasmid replication process by enzymes called topoisomerases. The supercoiled conformation can be maintained when both polynucleotide strands are intact, hence called covalently closed-circular (ccc) DNA. If one of the polynucleotide strands is broken, the double helix reverts ...
... plasmid DNA during the plasmid replication process by enzymes called topoisomerases. The supercoiled conformation can be maintained when both polynucleotide strands are intact, hence called covalently closed-circular (ccc) DNA. If one of the polynucleotide strands is broken, the double helix reverts ...
Lab IV: Recombinant DNA Analysis
... cells plated on agar-based media. Media additives such as antibiotics or X-gal can select for the presence of certain genes (and therefore certain fragments) within the transformed plasmid. Transformation is a rare event, and so a single colony on selective media is in all likelihood derived from a ...
... cells plated on agar-based media. Media additives such as antibiotics or X-gal can select for the presence of certain genes (and therefore certain fragments) within the transformed plasmid. Transformation is a rare event, and so a single colony on selective media is in all likelihood derived from a ...
Relationships between a new type IV secretion system and the icm
... When a sequence comparison of the L. pneumophila Lvh proteins with all the other known proteins of type IV secretion systems was performed, we found that from the seven conserved proteins (VirB4, -B7, -B8, -B9, -B10, -B11 and -D4) of type IV secretion systems, four of the L. pneumophila Lvh proteins ...
... When a sequence comparison of the L. pneumophila Lvh proteins with all the other known proteins of type IV secretion systems was performed, we found that from the seven conserved proteins (VirB4, -B7, -B8, -B9, -B10, -B11 and -D4) of type IV secretion systems, four of the L. pneumophila Lvh proteins ...
Protocol: Cloning of oligos for sgRNA (CRISPR) or
... types of pXPR vectors can be used: 1. pXPR_003 (or “lenti guide”) will only contain an sgRNA and is to be used in a cell line that already expresses Cas9. 2. pXPR_023 (also referred to as “lenti CRISPR” or “all-in-one”) already contains Cas9 sequence prior to cloning an sgRNA -For shRNA, pLKO vector ...
... types of pXPR vectors can be used: 1. pXPR_003 (or “lenti guide”) will only contain an sgRNA and is to be used in a cell line that already expresses Cas9. 2. pXPR_023 (also referred to as “lenti CRISPR” or “all-in-one”) already contains Cas9 sequence prior to cloning an sgRNA -For shRNA, pLKO vector ...
(S) tet Resistance Determinant Element Containing the Tetracycline
... initially identified in a multiresistant Listeria monocytogenes strain on a 37-kb conjugative plasmid, pIP811 (2). Subsequently tet(S) has been found on plasmid pK214 from Lactococcus lactis (8) and in the chromosome of Enterococcus faecalis (3). The tet(S) gene in L. lactis and L. monocytogenes is ...
... initially identified in a multiresistant Listeria monocytogenes strain on a 37-kb conjugative plasmid, pIP811 (2). Subsequently tet(S) has been found on plasmid pK214 from Lactococcus lactis (8) and in the chromosome of Enterococcus faecalis (3). The tet(S) gene in L. lactis and L. monocytogenes is ...
Functional analysis of plastid DNA replication origins in tobacco by
... and G; no participation in DNA replication has been ascribed to them up to now. (b) 322 bp downstream of oriA is a 852-bp DNA element termed NICE1, which has been found in extrachromosomal DNA circles in transformed tobacco chloroplasts, also in multimeric form (Staub and Maliga, 1994). It is border ...
... and G; no participation in DNA replication has been ascribed to them up to now. (b) 322 bp downstream of oriA is a 852-bp DNA element termed NICE1, which has been found in extrachromosomal DNA circles in transformed tobacco chloroplasts, also in multimeric form (Staub and Maliga, 1994). It is border ...
video slide
... – Thus the cDNA library is made of a set of genes that were transcribed in the starting cells and the new DNA strand that is produced is called complementary DNA or cDNA. – This cDNA represents only part of the genome – This is advantageous for; • Studying the genes responsible for specialized funct ...
... – Thus the cDNA library is made of a set of genes that were transcribed in the starting cells and the new DNA strand that is produced is called complementary DNA or cDNA. – This cDNA represents only part of the genome – This is advantageous for; • Studying the genes responsible for specialized funct ...
Molecular cloning, characterization, and homologous
... °C for 3 min. The final extension was performed for 10 min at 72 °C. Reaction mixtures were run in a 0.8% agarose gel. PCR products were extracted from the gel and ligated to pGEM-T Easy Vector (Promega). Ligation product was then transferred into the competent E. coli DH5α cells, which were prepare ...
... °C for 3 min. The final extension was performed for 10 min at 72 °C. Reaction mixtures were run in a 0.8% agarose gel. PCR products were extracted from the gel and ligated to pGEM-T Easy Vector (Promega). Ligation product was then transferred into the competent E. coli DH5α cells, which were prepare ...
Cell Structure and Function
... surface of bacteria • May stabilize membrane • Bacterium can vary structure to evade immune system • Can cause shock ...
... surface of bacteria • May stabilize membrane • Bacterium can vary structure to evade immune system • Can cause shock ...
![nature | methods Versatile P[acman] BAC libraries for transgenesis](http://s1.studyres.com/store/data/016819563_1-9796374085bf2dc123930427a979e4a9-300x300.png)
nature | methods Versatile P[acman] BAC libraries for transgenesis
... Two plasmid DNA preparation methods were used; these can be substituted. For CHORI322 clones, cultures were induced to high plasmid copy number with CopyControl™ Fosmid Autoinduction Solution (Epicentre) in LB (12.5 µg/ml Chl) for 17 hrs at 37°C. Plasmid was isolated using the PureLinkTM HiPure Plas ...
... Two plasmid DNA preparation methods were used; these can be substituted. For CHORI322 clones, cultures were induced to high plasmid copy number with CopyControl™ Fosmid Autoinduction Solution (Epicentre) in LB (12.5 µg/ml Chl) for 17 hrs at 37°C. Plasmid was isolated using the PureLinkTM HiPure Plas ...
Eukaryotic DNA Replication
... Any segment of DNA that has an origin should be able to replicate, so although plasmids are rare in eukaryotes, it may be possible to construct them by suitable manipulation in vivo. This has been accomplished in yeast, although not in higher eukaryotes. The discovery of yeast origins resulted ...
... Any segment of DNA that has an origin should be able to replicate, so although plasmids are rare in eukaryotes, it may be possible to construct them by suitable manipulation in vivo. This has been accomplished in yeast, although not in higher eukaryotes. The discovery of yeast origins resulted ...
DNA Scissors: Introduction to Restriction
... 2. Now separate the hydrogen bonds between the cut sites by cutting through the vertical lines. Separate the two pieces of DNA. Look at the new DNA ends produced by EcoRI. Are they sticky or blunt? Write EcoRI on the cut ends. Keep the cut fragments on your desk. 3. Repeat the procedure with strip 2 ...
... 2. Now separate the hydrogen bonds between the cut sites by cutting through the vertical lines. Separate the two pieces of DNA. Look at the new DNA ends produced by EcoRI. Are they sticky or blunt? Write EcoRI on the cut ends. Keep the cut fragments on your desk. 3. Repeat the procedure with strip 2 ...
iGenetics: A Molecular Approach, 3e (Russell/Bose)
... 32) Cloning vectors generally possess unique restriction sites and dominant selectable markers. Answer: TRUE Skill: Factual recall 33) A linear strand of DNA with two restriction enzyme cut sites will yield three fragments on digestion. Answer: TRUE Skill: Problem-solving 34) A circular DNA molecule ...
... 32) Cloning vectors generally possess unique restriction sites and dominant selectable markers. Answer: TRUE Skill: Factual recall 33) A linear strand of DNA with two restriction enzyme cut sites will yield three fragments on digestion. Answer: TRUE Skill: Problem-solving 34) A circular DNA molecule ...
Regulation of Bacterial Conjugation in Microaerobiosis by
... The virulence plasmid of Salmonella enterica (pSLT) is an F-like conjugative plasmid. High rates of pSLT transfer occur in the mammalian gut, a microaerobic environment. In this study, we describe genetic screens for host-encoded activators and repressors of the transfer operon (tra) of pSLT. We sho ...
... The virulence plasmid of Salmonella enterica (pSLT) is an F-like conjugative plasmid. High rates of pSLT transfer occur in the mammalian gut, a microaerobic environment. In this study, we describe genetic screens for host-encoded activators and repressors of the transfer operon (tra) of pSLT. We sho ...
Full-Text PDF
... the system of choice for studying herpesviruses. BAC vectors are based on the E. coli fertility factor (F-factor) replicon which is maintained as a circular supercoiled extrachromosomal single copy plasmid in the bacterial host [2,30,31]. BACs can accept inserts up to 300 Kb in length, which is suff ...
... the system of choice for studying herpesviruses. BAC vectors are based on the E. coli fertility factor (F-factor) replicon which is maintained as a circular supercoiled extrachromosomal single copy plasmid in the bacterial host [2,30,31]. BACs can accept inserts up to 300 Kb in length, which is suff ...
Prolonged organ retention and safety of plasmid DNA
... dosing of PEI/DNA complexes over 3 weeks did not alter the histology of major organs such as the liver, lung, and kidney. PEI/DNA complexes with PEI nitrogen to DNA phosphate ratio (N/P ratio) of 10:1 were prepared in 5% glucose using 25 kDa PEI (Aldrich, St Quentin, France). The N/P ratio of 10:1 w ...
... dosing of PEI/DNA complexes over 3 weeks did not alter the histology of major organs such as the liver, lung, and kidney. PEI/DNA complexes with PEI nitrogen to DNA phosphate ratio (N/P ratio) of 10:1 were prepared in 5% glucose using 25 kDa PEI (Aldrich, St Quentin, France). The N/P ratio of 10:1 w ...
Recombinant DNA Lesson - Ms. Guiotto Biology Class
... placed in a solution containing a ligase, recombination occurs at random. Many recombinations are possible, and a fraction of these contain the desired recombinant DNA. For example, if one of the fragments with complementary ends was a ...
... placed in a solution containing a ligase, recombination occurs at random. Many recombinations are possible, and a fraction of these contain the desired recombinant DNA. For example, if one of the fragments with complementary ends was a ...
Restriction Enzyme Digest and Plasmid mapping
... This excercise introduces you to some important principles of genetic engineering. Specifically, the functions of restriction enzymes and their use as molecular biology tools will be stressed. Using agarose gel electrophoresis, you will examine the digestion patterns and determine the sizes of unkno ...
... This excercise introduces you to some important principles of genetic engineering. Specifically, the functions of restriction enzymes and their use as molecular biology tools will be stressed. Using agarose gel electrophoresis, you will examine the digestion patterns and determine the sizes of unkno ...
Molecular analysis of an operon in Bacillus subtilis
... Sanger e t al. (1977) and synthetic oligonucleotide primers were used. ...
... Sanger e t al. (1977) and synthetic oligonucleotide primers were used. ...
12.3 How Is Biotechnology Used In Forensic Science?
... parts of genes from different organisms. • Large amounts of recombinant DNA can be grown in bacteria, viruses, or yeasts, and then transferred into other species. • Plants or animals that express DNA that has been modified or derived from other species are called ______________, or genetically modif ...
... parts of genes from different organisms. • Large amounts of recombinant DNA can be grown in bacteria, viruses, or yeasts, and then transferred into other species. • Plants or animals that express DNA that has been modified or derived from other species are called ______________, or genetically modif ...
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.