LG GCE-8520A DRIVER DETAILS:
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LG GCE-8520A DRIVER
DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation by biological means, e. The invention also relates to a procedure for obtaining these vectors as well as transgenic plants LG GCE-8520A them. In general, the vectors are known in the field of biotechnology and genetic manipulation. The vectors which are currently used most commonly for genetic transformation, and in particular in the field of plant biotechnology, present several disadvantages in their use, however. In actual fact, and notably for plant transgenesis, use has often been made of a vector called pBinl9 Frisch et al.
The nucleotide sequence of this binary plasmid LG GCE-8520A is entirely known. The problem, however, LG GCE-8520A that this plasmid is of Large size Moreover, the selection cassette of this plasmid is located near the right border of the T-DNA. Thus, when the T-DNA is broken after the selection cassette, it will not be possible to make any selection in order to retain solely the plants possessing the expression cassette of the desired gene. The expressions used in the description and the claims have the following meaning.
If a micro-organism or a recombinant cell culture is described as host of an "expression vector", LG GCE-8520A can also include extrachromosomal circular DNA such, as for example mitochondrial or chloropiast DNADNA which has been integrated into the host chromosome swhere the vector can be either replicated in a stable manner by the cells during mitosis as an autonomous structure, integrated into the genome of the host, or maintained in the nucleus or cytoplasm of the host. In this case, the "plasmid" is a molecule of autonomous circular DNA capable of replication in a cell.
If a micro-organism or recombinant cell culture is described as the host of an "expression" plasmid, this comprises both extrachromosomal circular DNA molecules and DNA which has been integrated into the host chromosome s. If the plasmid is maintained by a host cell, the plasmid is either LG GCE-8520A in a stable manner by the cells during mitosis as an autonomous structure, or integrated into the genome of the host; - "clean" means that the vector comprises only sequences that are indispensable for its functionality and carries a nucleic acid sequence which comprises only elements that are indispensable for the expression of the host cell; - "nucleic acid" means DNA or RNA ; - "nucleic acid sequence" means a single- or double-stranded oligomer or polymer of nucleotide bases read from the 5' end towards the 3' end, and comprises self-replicating plasmids, genes, DNA or RNA polymers, which may or may not be infectious, and DNA, or RNA, either functional or non-functional.
Such cassettes include at least one promoter and a transcription termination signal, and optionally other factors necessary or useful for the expression ; - "heterologous sequence" or "heterologous nucleic acid sequence" means a sequence originating from a source, or from a species, foreign to the environment thereof, or if it originates from LG GCE-8520A same environment, which has been modified relative to its original form.
The modification of the nucleic acid sequence can take place for example by treatment of the nucleic acid with a restriction enzyme in order to generate a nucleic acid fragment which can be operationally bound to a promoter. The modification can also take place by means of techniques such as directed mutagenesis; - "box" means a nucleic acid sequence to which a regulatory function LG GCE-8520A attributed: The percentage of sequence identity is calculated on the basis of a comparison window of at least 6 nucleotide bases.
The position which is given by a figure refers to the position of the start of the element in the nucleic acid sequence, in the direction of reading of the latter, i. The transgenic plants according to the present invention can have different levels of ploidy, and can notably be polyploid, diploid, and haploid; - "propagule" means a cluster or association of plant cells, which may or may not be structured, allowing the regeneration of a whole plant, for example expiants, calli, stems, leaves, roots, cuttings, and even seeds.
The applicant of the present invention has succeeded, surprisingly, in producing clean synthetic vectors, in particular binary plasmids, of completely known nucleotide sequence, of small size, allowing the aforesaid disadvantages to be alleviated, and notably presenting a high replication rate relative to the existing vectors most commonly used. Furthermore, the applicant has succeeded at the same time in producing a range of vectors in such a way as to be able to choose the one which it is convenient to use according to the application envisaged and the environment of its use, and thus in such a way as to be able to better control the rate of expression of a gene to be expressed, coding for a polypeptide to be produced. In addition, in some of the vectors according to the invention, each of the functional elements or components can be isolated by simple enzymatic digestion.
An object of the present invention is therefore a clean synthetic vector containing only the elements indispensable to its functionality and to the transgenesis of a cell, and notably of a plant cell. According to a preferred embodiment, the vector comprises, as elements which are indispensable to its LG GCE-8520A and to the transgenesis of a cell, and which are operationally bound: According to a LG GCE-8520A preferred embodiment the vector comprises the nucleic acid sequence identified by the number SEQ. Still more preferably, the vector consists of a single plasmid pMRT whose nucleic acid sequence is identified by the number SEQ.
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According to another preferred embodiment, the vector includes at least one nucleic acid sequence coding for a second origin of replication, preferably LG GCE-8520A on of Escherichia coli, and more preferably an on ColEI. More preferably, the vector comprises the nucleic acid sequence identified by the number SEQ.
ID02, and still more preferably, the vector consists of a single plasmid pMRT whose nucleic acid sequence is identified by the number SEQ. According to a preferred embodiment, the vector also includes a nucleic acid sequence coding for a T-DNA comprising a right border RB and a left border LB, allowing the vector to act as a binary plasmid. According to another preferred embodiment, the vector also includes a nucleic acid sequence coding for at least one expression promoter and at least one transcription terminator situated between the left border LB and the right border LG GCE-8520A of the T-DNA.
More preferably, the expression promoter is chosen from the group consisting of the constitutive promoters, the LG GCE-8520A promoters, the specific promoters, and preferably chosen from the plant expression promoters. Preferably, the expression terminator is chosen from the functional LG GCE-8520A in a plant cell, and is preferably a 35S or nos terminator. More preferably still, the nucleic acid sequence coding for the selection agent is situated near the left border of the T-DNA.
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According to another preferred embodiment of the present invention, the vector includes at least one expression cassette comprising an expression-promoting nucleic acid sequence operationally bound to a nucleic acid sequence to be expressed, coding for a polypeptide to be produced, itself bound to a transcription termination nucleic acid sequence. Examples of such proteins are for example the insulins, interferons, gastric, pancreatic or biliary lipases, elastases, antiproteases such as alpha-1 antitrypsin, structural proteins such as collagen, transferrins such as lactoferrin, proteins derived from blood, such as haemoglobin, human albumin and the blood cofactors, and antioxidants such as superoxide dismutase, According to a particularly preferred embodiment of the invention, the vector is presented in the form of a binary, linear or circular plasmid, chosen from the group consisting of the nucleic acid sequences identified by the numbers SEQ.
According to a particularly advantageous LG GCE-8520A, each functional component of the vector can be cleaved independently of the other components. NOTE: You are now downloading LG GCRA firmware a.
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