Biochemistry of P element transposase, the mechanism of transposition and P element-related THAP 9 genes in humans and zebrafish. The 87kD P element-encoded transposase protein is required to catalyze P element transposition and belongs to a large polynucleotidyl transferase superfamily, that includes RNaseH, RuvC, retroviral integrases, transposases and the argonaute proteins. Biochemical studies using the purified protein revealed that guanosine triphosphate (GTP) is an essential cofactor for the reaction. Current studies involve the use of imaging methods, including atomic force microscopy (AFM), electron microscopy (EM) and total internal reflection fluorescence microscopy (TIRFM) to understand the role(s) that GTP plays in transposition, the detailed reaction pathway and how this cofactor modulates the assembly and activity of transposase on DNA. We are also interested in using X-ray crystallography to study P element transposase bound to DNA, and initially in collaboration with James ...
This chapter describes the family of Tc1/ mariner elements, their mechanism of transposition, and the regulation of transposition. It also talks about a few applications of this family of transposable element in forward and reverse genetics. The transposase proteins contain the typical DDE or DDD motif found in most transposases and integrases. Excision of P elements in Drosophila melanogaster has been used as a trigger to initiate introduction of new sequences into the original P element-containing site; it has been shown also for Tc1 that an ectopic transgenic template (marked by polymorphisms) can be used by the repair process. For several members of the Tc1/mariner transposase family a smaller truncated version of the transposase was seen. Possibly it is the way of reconstructing Sleeping Beauty transposase that is responsible for generating a protein that is more active than any element found in nature, where it is not in the best interest of the parasite transposable element to encode a
Bacterial insertion sequences are the simplest form of autonomous mobile DNA. It is unknown whether they need to have beneficial effects to infect and persist in bacterial populations, or whether horizontal gene transfer suffices for their persistence. We address this question by using branching process models to investigate the critical, early phase of an insertion sequence infection. We find that the probability of a successful infection is low and depends linearly on the difference between the rate of horizontal gene transfer and the fitness cost of the insertion sequences. Our models show that the median time to extinction of an insertion sequence that dies out is very short, while the median time for a successful infection to reach a modest population size is very long. We conclude that horizontal gene transfer is strong enough to allow the persistence of insertion sequences, although infection is an erratic and slow process. ...
TY - JOUR. T1 - (+)-CC-1065 as a structural probe of Mu transposase-induced bending of DNA. T2 - Overcoming limitations of hydroxyl-radical footprinting. AU - Ding, Zhi Ming. AU - Harshey, Rasika M.. AU - Hurley, Laurence. PY - 1993/9/11. Y1 - 1993/9/11. N2 - Phage Mu transposase (A-protein) is primarily responsible for transposition of the Mu genome. The protein binds to six att sites, three at each end of Mu DNA. At most att sites interaction of a protein monomer with DNA is seen to occur over three minor and two consecutive major grooves and to result in bending up to about 90°. To probe the directionality and locus of these A-protein-induced bends, we have used the antitumor antibiotic (+)-CC-1065 as a structural probe. As a consequence of binding within the minor groove, (+)-CC-1065 is able to alkylate N3 of adenine in a sequence selective manner. This selectivity is partially determined by conformational flexibility of the DNA sequence, and the covalent adduct has a bent DNA structure in ...
ISfinder (www-is.biotoul.fr) is a dedicated database for bacterial insertion sequences (ISs). It has superseded the Stanford reference center. One of its functions is to assign IS names and to provide a focal point for a coherent nomenclature. It is also the repository for ISs. Each new IS is indexe …
Cell culture and transposition assay HEK 293 cells had been maintained in MEMa medium supplemented with 10% FBS, 100 units ml penicillin, and a hundred ug mL streptomycin. The specifics for your transposition assays had been described pre viously. Inhibitors,Modulators,Libraries Exercise assay in the piggyBac transposase A equivalent procedure as thorough previously was employed to co transfect a hundred ng of piggyBac donor, with various quantity of the piggyBac helper, pCMV Myc piggyBac, ranging from 0 300 ng into 1. 2 105 of HEK 293 cells. pcNDA3. 1NEO, an empty vector used in our prior examine, was employed to best the total level of DNA transfected to 400 ng. Each and every trans fection condition was completed in triplicate. Twenty 4 hours just after transfection, one fifth of transfected cells were subjected to transposition assay.. The remaining transfected cells in triplicate had been pooled and grew in the 35 mm plate for a further twenty four hours prior to staying subjected to ...
In this study, first we showed that the Tol2 transposon system is a useful technique in generating stable transfected primary culture cells and, second, we demonstrated that the Tol2 transposon system is applicable to the study of circadian clock oscillations.. The Tol2 transposon was originally discovered from Medaka fish (Orzyias latipes) [12]. An active autonomous member of Tol2 was first identified by the analysis using zebrafish embryos [13]. Since then, the Tol2 transposon system has been mainly used for random insertion mutagenesis and transgene in zebrafish [14]. Although recent reports have indicated that the transposon systems such as piggyBac and SleepingBeauty in addition to Tol2 are also active in mammalian cells [15, 16], few studies have been reported that utilized the Tol2 system for transfection to mammalian primary culture cells. In the present study, we showed that the Tol2 transposon system is a useful tool in generating stable transfected primary culture cells such as MEF ...
A versatile genetic method for identifying and cloning Drosophila melanogaster genes affecting any recognizable phenotype is described. Strains are constructed in which the insertion of a single P transposable element has caused a new mutation, greatly simplifying the genetic and molecular analysis of the affected gene. Mutagenesis is initiated by crossing two strains, each of which contains a specially designed P element. One element (jumpstarter), encoding P element transposase, efficiently mobilizes the second nonautonomous transposon (mutator), whose structure facilitates selection and cloning of new insertion mutations. Random mutator transpositions are captured in individual stocks that no longer contain jumpstarter, where they remain stable. This method was used to construct 1300 single P element insertion stocks which were then screened for recessive mutations. A library of single-element insertion strains will allow the structure and function of Drosophila genes to be readily ...
Transposase is an enzyme that binds to the end of a transposon and catalyzes the movement of the transposon to another part of the genome by a cut and paste mechanism or a replicative transposition mechanism. The word "transposase" was first coined by the individuals who cloned the enzyme required for transposition of the Tn3 transposon. The existence of transposons was postulated in the late 1940s by Barbara McClintock, who was studying the inheritance of maize, but the actual molecular basis for transposition was described by later groups. McClintock discovered that pieces of the chromosomes changed their position, jumping from one chromosome to another. The repositioning of these transposons (which coded for color) allowed other genes for pigment to be expressed. Transposition in maize causes changes in color; however, in other organisms, such as bacteria, it can cause antibiotic resistance. Transposition is also important in creating genetic diversity within species and adaptability to ...
Until recently, a transposon popularly known as Sleeping Beauty was thought to be the most efficient in inserting genes into mammalian systems. However, a drawback to Sleeping Beauty is its inability to be modified to overcome concerns about placing therapeutic genes safely within the genome. Moisyadi and Kaminski compared a number of transposons and found that piggyBac is much more efficient at inserting transgenes into the genome of several human lines. It is also amenable to molecular alteration so that it can potentially insert a therapeutic gene into a safe area of the genome. In fact, the piggyBac transposase enzyme that performs the insertion of the transposon into DNA can now be modified to attempt placing the transgenes into pre-designated regions of the genome - targeted insertion ...
Transposase activity was thought to be extinct in humans because DNA movement can be deleterious in higher organisms, resulting in genomic instability and perha...
Interestingly there is no rule such as the higher evolved an organism the more transposable elements. Although we find more transposable elements in higher evolved organisms this rule can not be maintained if compared single species as frogs and humans.. Transposons most often code for transposase the enzyme responsible for the transposon dislocation. As Transposons are so common in a genome it is not surprising that transposases are the most common genes in a genome [2].. ...
Previous studies on the genetic environment of blaKPC have identified several ORFs encoding putative transposases located upstream and downstream of the blaKPC genes. In the present work, we have further characterized the genetic environment of the blaKPC gene by detailed analysis of a 22-kb insert derived from the natural plasmid pNYC-1 containing the blaKPC gene from K. pneumoniae isolate YC from Paris but with a U.S. origin (16) and by analysis of blaKPC-containing natural plasmids from isolates from Greece (6) and Colombia (28, 29) and blaKPC-containing sequences available in the GenBank database. We were able to identify a novel Tn3-based transposon, Tn4401, which is at the origin of blaKPC-like gene acquisition and dissemination. In addition to the tnpA transposase, Tn4401 possesses the resolvase tnpR, the blaKPC gene, and two ISs, ISKpn6 and ISkpn7. These ISs must have inserted into the parental transposon, since both ISs are flanked by target site duplications, signaling a recent ...
Transposons have been used in invertebrates for transgenesis and insertional mutagens in genetic screens. We tested a functional transposon called Sleeping Beauty in the one-cell mouse embryo. In this report, we describe experiments in which transposon vectors were injected into one-cell mouse embryos with mRNA expressing the SB10 transposase enzyme. Molecular evidence of transposition was obtained by cloning of insertion sites from multiple transgenic mice produced by SB10 mRNA/transposon coinjection. We also demonstrate germ-line transmission and expression from transposed elements. This technique has promise as a germ-line transgenesis method in other vertebrate species and for insertional mutagenesis in the mouse ...
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At the beginning of this week we spoke to Dr. Michael Hynes, who was able to give us E. coli SM10 and SM17-1 cells containing the plasmid pOT182. This plasmid contains an E. coli origin of replication, allowing it to act as a suicide vector when transferred to a different bacterial species. pOT182 contains a Tn5 transposon element containing a promotorless lacZ gene, genes for tetracycline resistance as well as a beta-lactamase, transposase and an E. coli origin of replication. These elements are bordered by insertion element sequences which are recognised by the transposase. When transferred to a different host through conjugation, the plasmid itself can no longer replicate. The transposase however can recognise and transfer the sequence between the insertion elements in a cut-and-paste fashion randomly into the genome. In this fashion, the tetracycline and beta-lactam resistant traits would only persist in cells in which the transposon has jumped into the genome, allowing these antibiotics to ...
The transposable element (TE), Tn5, is a conservative transposon that is able to insert a segment of genes bordered by specific 19bp insertion sequences (IS) from one part of the genome (e.g. plasmid vector) randomly to another location, such as the chromosome (Reznikoff, 2008). The transposition event is catalyzed by a transposase enzyme encoded by Tnp gene included in the TE. By inserting a vector construct containing the TE with selectable markers (such as tetracyclin resistance and lacZ) into an organism with a desirable phenotype, we can find out what genetic elements (e.g. genes and promoters) are responsible for that particular function. This can happen via a random insertion of a TE containing a promoterless reporter gene downstream of promoter elements that creates a transcriptional fusion, providing activity in response to specific environmental stimuli. Another advantage of using a transposon approach is that it creates a saturating library of mutants where all possible genetic ...
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These reference sequences exist independently of genome builds. Explain. These reference sequences are curated independently of the genome annotation cycle, so their versions may not match the RefSeq versions in the current genome build. Identify version mismatches by comparing the version of the RefSeq in this section to the one reported in Genomic regions, transcripts, and products above. ...
Immusofts mission is to develop a breakthrough autologous cell therapy platform for treating a variety of human diseases through our proprietary Immune System Programming (ISP™) technology. ISP™ technology can effectively re-program a patients own cells to become miniature drug factories in the body. The technology was designed to address current challenges faced with the production and delivery of conventional protein therapeutic drugs (biologics). The ISP™ platform enables safe insertion of genes encoding the correct human homolog of a missing or defective protein(s) into a patients immune cells using the Sleeping Beauty (SB) transposon system - a non-viral vector ...
Mariner: Mariner, any of a series of unmanned U.S. space probes sent to the vicinities of Venus, Mars, and Mercury. Mariner 1 (launched July 22, 1962) was intended to fly by Venus, but it was destroyed shortly after liftoff when it veered off course. Mariners 2 (launched Aug. 27, 1962) and 5 (launched June
The two-element transposon constructs, utilizing either Ac/Ds or Spm/dSpm, allow random tagging of genes in heterologous model species, but are inadequate for directed tagging of specific alleles of agronomic importance. We propose the use of Ac/Ds in conjunction with Spm/dSpm to develop a four-element system for directed tagging of crop-specific alleles. The four-element based construct would include both Ds and dSpm along with relevant marker genes and would function in two steps. In the first step dSpm(Ds) stocks (a minimum of two) would be crossed to a line containing transposases of Spm and unlinked integrations would be selected from segregating population by the use of a negative selection marker to develop stocks representing integration of dSpm(Ds) at a large number of locations in the genome. Selections would be made for a line in which dSpm(Ds) shows partial or complete linkage to the allele of interest. In the second step selected line would be crossed to a line containing Ac ...
Exploring Life - Honors Biology Chapter 9 Cell Cycle, Mitosis & Meiosis. When you click Submit (at the bottom of this web page) it will save your work on the server for me. If you get a proxy error hit -refresh- until you get a Thanks. You might want to actually write your answers in Word (so you can spell check them) and then cut-and-paste into the boxes. That way, if the computer misplaces your work you can just re-paste the answers.. Since we may work on this over more than one day, I will keep track of what you have done. Feel free to click submit as you work to save your work to the server. DO NOT JUST LOOK FOR THE ANSWERS YOU LL MISS LOTS OF IMPORTANT STUFF. THE QUESTIONS ARE JUST TO KEEP YOU HONEST. TAB OR USE THE MOUSE Between the fields..... Answer the questions as best you can. The boxes will expand as you type if you need more space. (Dont worry if it is a one word answer. The box size is standard.). Since you will be working in groups, put all the names of the people in the ...
Actually, Game is like any tool. You can use it in a moral or immoral way. One man I recommended it to, said that he could not use it.. Another man said that the only way he would use it, is if he discussed it with his wife and she approved. Here is a cut-and-paste from a couple of comments I made over at manboobz:. http://manboobz.com/2011/06/22/arms-and-the-mens-rights-movement/comment-page-9/#comment-33758. AndrewV , June 27, 2011 at 1:23 am denelian , June 27, 2011 at 12:15 am. â maybe i should go find a PUA website and read more. any suggestions?â I would recommend this guy Roissy ...
Applied Statistics Assignment 1 Due Fri 29 March 2013 Instructions 1. Cut-and-paste this document into word (or equivalent) 2. Show working where possible...
Dont look now, but the 2013 Seattle Mariners can hit. I know, I know, youre waiting for the punchline, right? After all, were talking about the Mariners...
Felix Hernandez dominated the Yankees in their own ballpark again, pitching his third shutout of the season to lead the Seattle Mariners to a 1-0 win on a...
Thanks also to our invaluable volunteers: Joanne Ball, Carole Coffey, Mary Gorman, Barbara Leavitt, Susan Ovenden, Bill Souden, Beverly Westerveld and Ed Bamford.
One of the questions on the survey at the feed (results to be posted sometime, I promise) was which contract signed this offseason is the worst for the signing
Bartolo Colon points as he retires Seattle's Yuniesky Betancourt with two runners in scoring position for the third out of the 7th inning during Sox 2-1 win.
Transposition allows movement of a defined stretch of DNA, a transposon, from one DNA location to another. This process is required for the life-cycles of many viruses, from bacteriophage Mu to HIV; it is spreading antibiotic resistances between bacterial populations; and it is responsible for spontaneous mutations in all the kingdoms of life. Transposition is mediated by a protein, the transposase, encoded by the transposon. DNA sequence signals at the two ends of the transposon activate assembly of a transpososome: a complex that include multiple copies of transposase plus both transposon ends. Transpososome assembly, in turn, activates the DNA cleavage and joining reactions required for transposition. This thesis explores aspects of interactions between one transposase, MuA, and the ends of its transposon DNA, the genome of bacteriophage Mu. The first chapter provides an overview of Mu transposition, with special emphasis on the transpososome. The second chapter shows that in the absence of ...
We describe discovery in |i|Beta vulgaris|/i| L. of |i|Coe1|/i|, a DNA transposase gene within putative long terminal repeats (LTRs), and other retrotransposon-like features including both a retroviral-like hypothetical gene and an Rvt2-domain reverse transcriptase pseudogene. The central DNA transposase gene encodes, in eight exons, a predicted |?bhlt ?|160-KDa|?ehlt ?| protein producing BLAST alignments with |i|En/Spm|/i|-type transposons. Except for a stop signal, another ORF encodes a |i|Ty1-copia|/i|-like reverse transcriptase with amino acid sequence domain YVDDIIL. Outside apparent LTRs, an 8-mer nucleotide sequence motif CACTATAA, near or within inverted repeat sequences, is hypothetical extreme termini. A genome scan of |i|Arabidopsis thaliana|/i| found another example of a |i|Tnp2|/i|-domain transposase gene within an apparent LTR-retrotransposon on chromosome 4.
Transposons are mobile DNA segments that can disrupt gene function by inserting in or near genes. Here, we show that insertional mutagenesis by the PiggyBac transposon can be used for cancer gene discovery in mice. PiggyBac transposition in genetically engineered transposon-transposase mice induced cancers whose type (hematopoietic versus solid) and latency were dependent on the regulatory elements introduced into transposons. Analysis of 63 hematopoietic tumors revealed that PiggyBac is capable of genome-wide mutagenesis. The Piggybac screen uncovered many cancer genes not identified in previous retroviral or Sleeping Beauty transposon screens, including Spic, which encodes a PU.1-related transcription factor, and Hdac7, a histone deacetylase gene. PiggyBac and Sleeping Beauty have different integration preferences. To maximize the utility of the tool, we engineered 20 mouse lines to be compatible with both transposases in constitutive, tissue-, or temporal-specific mutagenesis. Mice with ...
Transposable elements have emerged as a promising candidate for human non-viral gene-therapy. The Tc1/mariner transposon Sleeping Beauty is to date one of the most efficient transposons in mammals. Sleeping Beauty transposase has so far mostly been delivered to cells via a DNA source. This might cause spontaneous integration of the transposase gene and cause fatal damage to the affected cell. Hence, it would be advantageous to employ a non-genetic source for the transposase. We here show that a novel Cell-penetrating peptide, M918, has the ability to facilitate cellular delivery of both the transposase Sleeping Beauty as a protein and a transposon donor-plasmid carrying an antibiotic resistance gene in vitro. The technique is a simple and straightforward one-step method that might render a safe and efficient delivery platform for Sleeping Beauty mediated gene therapy.. ...
Random gene inactivation used to identify cellular functions associated with virulence and survival of Brucella spp has relied heavily upon the use of the transposon Tn5 that integrates at G/C base pairs. Transposons of the mariner family do not require species-specific host factors for efficient transposition, integrate nonspecifically at T/A base pairs, and, at a minimum, provide an alternative approach for gene discovery. In this study, plasmid vector pSC189, containing both the hyperactive transposase C9 and transposon terminal inverted repeats flanking a kanamycin resistance gene, were used to deliver Himar1 transposable element into the B. melitensis genome. Conjugation was performed efficiently and rapidly in less than one generation in order to minimize the formation of siblings while assuring the highest level of genome coverage. Although previously identified groups or classes of genes required for virulence and survival were represented in the screen, additional novel identifications were
1BCM: Structure of the bacteriophage Mu transposase core: a common structural motif for DNA transposition and retroviral integration.
PiggyBac Transposable Element Derived 5 is an enzyme that in humans is encoded by the PGBD5 gene.[1] PGBD5 is a DNA transposase related to the ancient PiggyBac transposase first identified in the cabbage looper moth, Trichoplusia ni.[2] The gene is believed to have been domesticated over 500 million years ago in the common ancestor of cephalochordates and vertebrates.[3] The putative catalytic triad of the protein composed of three aspartic acid residues is conserved among PGBD5-like genes through evolution,[4], and is distinct from other PiggyBac-like genes.[3] PGBD5 has been shown to be able to transpose DNA in a sequence-specific, cut-and-paste fashion.[4] PGBD5 has also been proposed to mediate site-specific DNA rearrangements in human tumors.[5] ...
We have isolated and characterized deletions arising within a P transposon, P[hswa], in the presence of P transposase. P[hswa] carries white-apricot (wa) sequences, including a complete copia element, under the control of an hsp70 promoter, and resembles the original wa allele in eye color phenotype. In the presence of P transposase, P[hswa] shows a high overall rate (approximately 3%) of germline mutations that result in increased eye pigmentation. Of 234 derivatives of P[hswa] with greatly increased eye pigmentation, at least 205 carried deletions within copia. Of these, 201 were precise deletions between the directly repeated 276-nucleotide copia long terminal repeats (LTRs), and four were unique deletions. High rates of transposase-induced precise deletion were observed within another P transposon carrying unrelated 599 nucleotide repeats (yeast 2 mu FLP; recombinase target sites) separated by 5.7 kb. Our observation that P element-mediated deletion formation occurs preferentially between ...
E. coli ClpX is a member of the Clp/Hsp100 family of ATPases that remodel multi-component complexes and facilitate ATP-dependent protein degradation. Protein remodelers alter the biological activity of their substrates, typically by changing the quaternary structure of their target proteins. ClpX remodels protein-DNA complexes, termed transpososomes, made during recombination of the phage Mu. When recombination is complete, the core four-subunit transpososome complex does not spontaneously release the DNA; transposase remains so stably bound that subsequent replication of the Mu genome is inhibited. To understand how ClpX releases the replication block without destroying the transpososomes, we characterized the mechanism and products of transpososome remodeling. To better understand the mechanism ClpX uses to facilitate remodeling, I first participated in a collaborative project that defined major biochemical reaction steps involved in protein degradation by ClpX and its associated peptidase ...
Both classes of transposable elements (DNA and RNA) are tightly regulated at the transcriptional level leading to the inactivation of transposition via epigenetic mechanisms. Due to the high copies number of these elements, the hypothesis has emerged that their regulation can coordinate a regulatory network of genes. Herein, we investigated whether transposition regulation of HsMar1, a human DNA transposon, differs in presence or absence of endogenous HsMar1 copies. In the case where HsMar1 transposition is regulated, the number of repetitive DNA sequences issued by HsMar1 and distributed in the human genome makes HsMar1 a good candidate to regulate neighboring gene expression by epigenetic mechanisms. A recombinant active HsMar1 copy was inserted in HeLa (human) and CHO (hamster) cells and its genomic excision monitored. We show that HsMar1 excision is blocked in HeLa cells, whereas CHO cells are competent to promote HsMar1 excision. We demonstrate that de novo HsMar1 insertions in HeLa cells (human)
Background: Tangle analysis has been applied successfully to study proteins which bind two segments of DNA and can knot and link circular DNA. We show how tangle analysis can be extended to model any stable protein-DNA complex. -- Results: We discuss a computational method for finding the topological conformation of DNA bound within a protein complex. We use an elementary invariant from knot theory called colorability to encode and search for possible DNA conformations. We apply this method to analyze the experimental results of Pathania, Jayaram, and Harshey (Cell 2002). We show that the only topological DNA conformation bound by Mu transposase which is biologically likely is the five crossing solution found by Pathania et al (although other possibilities are discussed). -- Conclusion: Our algorithm can be used to analyze the results of the experimental technique described in Pathania et al in order to determine the topological conformation of DNA bound within a stable protein-DNA complex ...
Drosophila P element mobilization is subject to a complex array of regulatory mechanisms. A fruitful approach to study them is the use of insertion mutations whose expression is influenced by P regulation. In the present report, it is shown that P element somatic products may influence the expression of an unrelated gene inserted in a P transposon. The P[wdl9.3]19De transgene carriers an in vitro modified white gene harboring a duplication of the 5 regulatory sequences. Expression of this transgene is repressed in a P background. No maternal effect is detected and repression can be relieved as soon as P chromosomes are replaced by M ones. The amplitude of repression is correlated to the P transposase activity of the individuals examined. Repression appears to be exerted by somatic products of complete autonomous P elements or of in vitro modified P elements lacking the capacity to express the fourth P exon. The P repression of P[wdl9.3]19DE is strongly dependent on the insertion site of this ...
Transposable elements are endogenous DNA sequences able to integrate into and multiply within genomes. They constitute a major source of genetic innovations, as they can not only rearrange genomes but also spread ready-to-use regulatory sequences able to modify host gene expression, and even can give birth to new host genes. As their evolutionary success depends on their vertical transmission, transposable elements are intrinsically linked to reproduction. In organisms with sexual reproduction, this implies that transposable elements have to manifest their transpositional activity in germ cells or their progenitors. The control of sexual development and function can be very versatile, and several studies have demonstrated the implication of transposable elements in the evolution of sex. In this review, we report the functional and evolutionary relationships between transposable elements and sexual reproduction in animals. In particular, we highlight how transposable elements can influence expression of
A Genetic Screen Using the PiggyBac Transposon in Haploid Cells Identifies Parp1 as a Mediator of Olaparib Toxicity. . Biblioteca virtual para leer y descargar libros, documentos, trabajos y tesis universitarias en PDF. Material universiario, documentación y tareas realizadas por universitarios en nuestra biblioteca. Para descargar gratis y para leer online.
Mutator-like transposable elements (MULEs) are widespread across fungi, plants and animals. Most of the research of MULEs has focused on plant where they are discovered and have significant impact on genome structure. Despite being widespread, only a few active MULEs have been identified, meanwhile, the transposition mechanism of the MULEs is previously unknown. Pack-MULEs are able to capture and amplify genes or gene fragments on a large scale, and a subset of plant Pack-MULEs have been shown to be likely playing functional roles in regulating gene expression and providing novel coding capacities. However, the presence of Pack-MULEs in non-plant species has not been reported.. In this study we report that Muta1 identified from the mosquito Aedes aegypti is capable of excision and reinsertion in a yeast transposition assay, element reinsertion generated either 8 bp or 9 bp target site duplications (TSDs) with no apparent sequence preference. Mutagenesis analysis revealed the importance of ...
1) a small deletion occurs in the transposase gene of an IS element and plasmid is integrated , 2) a small deletion occurs in the transposase gene of an IS element , 3) two IS elements integrate into a chromosome with only a small distance separating them , 4) an IS element integrates with another IS element with the help of a plasmid
misc{7861784, abstract = {DNA transposons are a class of mobile genetic elements that can autonomously move from one genomic location to another. They are powerful drivers of genetic change and have played a significant role in the evolution of many genomes. One such transposable element, IS608 from Helicobacter pylori employs a unique mechanism of transposition as it transposes in a single-stranded DNA form and inserts specifically 3 of a specific tetranucleotide sequence (Kersulyte et al., 2002; Guynet et al., 2008). Previous structural (Ronning et al., 2005; Barabas et al, 2008) and biochemical studies (Ton-Hoang et al., 2005) of IS608, revealed that the element chooses its integration site specifically via base-pairing between the transposon and the target DNA. This unique feature allowed re-directing transposon integration to various four-nucleotide sequences by simply modifying the transposon DNA sequence. A key feature of the retargeting strategy was that, unlike the previous attempts to ...
pUT mini-Tn5 System. The mini-Tn5 are mobile elements able to transpose to the chromosome from a delivery plasmid, by means of the transposase activity encoded by the tnp* gene, which is present in cis in the plasmid but external to the mini-Tn5 element. pUT mini-Tn5 vectors can be used to insert any DNA fragment in the bacterial chromosome, by cloning it within the mini-Tn5 region. Due to the loss of the tnp* gene after insertion, minitransposons are stably integrated into the chromosome and inherited. The different pUTmini-Tn5 vectors can then be used for repeated insertion events and therefore multiple insertions in the same strain are only limited by the availability of distinct selection markers.. Advantages:. ...
The mini-Tn5 are mobile elements able to transpose to the chromosome from a delivery plasmid, by means of the transposase activity encoded by the tnp* gene, which is present in cis in the plasmid but external to the mini-Tn5 element. pUT mini-Tn5 vectors can be used to insert any DNA fragment in the bacterial chromosome, by cloning it within the mini-Tn5 region. Due to the loss of the tnp* gene after insertion, minitransposons are stably integrated into the chromosome and inherited. The different pUTmini-Tn5 vectors can then be used for repeated insertion events and therefore multiple insertions in the same strain are only limited by the availability of distinct selection markers.. Advantages:. ...
Identification of Genes Encoding Exported Mycobacterium tuberculosis Proteins Using a Tn552′phoA In Vitro Transposition System: Secreted and cell envelope-assoc