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    Gateway(r) cloning is a high throughput technology based on site specific recombination reaction between different attachment sites. BP reaction is a reaction between an Expression Clone (or an attB-flanked PC R product) and Donor Vector containing attP sites to create an Entry Vector. LR reaction is between an Entry Vector (containing attR) and Destination Vector (containing expression C lone) this vector recombines with a Donor Vector to

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    Gateway(r) cloning is a high throughput technology based on site specific recombination reaction between different attachment sites. BP reaction is a reaction between an Expression Clone (or an attB-flanked PC R product) and Donor Vector containing attP sites to create an Entry Vector. LR reaction is between an Entry Vector (containing attR) and Destination Vector (containing expression C lone) this vector recombines with a Donor Vector to

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    33 views1 page

    Design Gateway Cloning Experiments

    Uploaded by

    sudhanshudwi
    Gateway(r) cloning is a high throughput technology based on site specific recombination reaction between different attachment sites. BP reaction is a reaction between an Expression Clone (or an attB-flanked PC R product) and Donor Vector containing attP sites to create an Entry Vector. LR reaction is between an Entry Vector (containing attR) and Destination Vector (containing expression C lone) this vector recombines with a Donor Vector to

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    Attribution Non-Commercial (BY-NC)
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    of 1

    6/6/13

    Design Gateway Cloning Experiments

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    Home >> Products >> SimVector >> GatewayC loning Glossary Terms

    SimVector
    Gateway Cloning
    Cloning: A process for obtaining genetically identical group of genes, cells or organisms. Gateway CloningCloning: A process for obtaining genetically identical group of genes, cells or organisms. Gateway Cloning: Gateway cloning is a high throughput technology based on site specific recombination reaction between different attachment sites, such as attP, phage attachment site; attB, bacterial attachment site; attL, left attachment junction and attR, right attachment junction. Using this technology, more than one gene can be transferred in one or more vectors, maintaining the reading frame and orientation. BP Reaction: It is the first recombination reaction of Gateway cloning. It is a reaction between an expression clone (or an attB-flanked PC R product) and donor vector containing attP sites to create an entry clone (PC R fragment with att B sites + Donor vector Entry C lone). LR Reaction: Is a recombination reaction between an entry clone (containing attL) and a destination vector (containing attR), mediated by a host of recombination proteins to generate an expression clone (Entry C lone + Destination Vector Expression C lone). Expression Clone/Vector : The term is generally used for a PC R amplified gene tagged with attB sites used in Gateway BP reaction. This vector recombines with a donor vector to generate an entry vector. Donor Vector : It is a vector used in Gateway system which recombines with an expression vector. The process is called a BP reaction and generates an entry vector. Entry Vector : The recombinant vector resulting from a Gateway BP reaction contains the cloned PC R amplified product. Technicals
    Gate way C loning Plasm id Drawing Plasm id Maps R e striction C loning R e striction Enzym e Analysis TA C loning O R F Se arch Ve ctorNTI

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    Destination Vector : A vector carrying a ccdB gene and an antibiotic resistance gene which are replaced by the gene of interest. The gene of interest is acquired from an entry vector. This process of transfer from an entry clone to a destination vector occurs in the LR reaction of a Gateway system generating an expression clone. att: It stands for attachment site generally present in phage viruses and bacteria. attB: Stands for bacterial attachment site which helps in site specific recombination and always attaches with the PC R product. It consist of four homologous core sequence of ~25 bp each, attB1 AC AAGTTTGTAC AAAAAAGC AGGC T attB2 AC C C AGC TTTC TTGTAC AAAGTGGT attB3 AC AAC TTTGTATAATAAAGTTG attB4 AC AAC TTTGTATAGAAAAGTTG SimVector Home SimVector Features

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