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OGS568

Sigma-Aldrich

PSF-CMV-MUKAPPA - MOUSE KAPPA LIGHT CHAIN ANTIBODY PLASMID

plasmid vector for molecular cloning

同義詞:

cloning vector, expression vector, molecular cloning vector, plasmid, plasmid vector, snapfast vector, vector

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About This Item

分類程式碼代碼:
12352200
NACRES:
NA.85

形狀

buffered aqueous solution

分子量

size 4576 bp

菌種選擇

kanamycin

複製起點

pUC (500 copies)

肽切割

no cleavage

啟動子

Promoter name: CMV
Promoter activity: constitutive
Promoter type: mammalian

報告基因

none

運輸包裝

ambient

儲存溫度

−20°C

一般說明

PSF-CMV-MUKAPPA – mouse kappa light chain antibody plasmid is a murine kappa light chain constant region expression plasmid. Variable antibody fragments can be fused in this plasmid to create full length antibody light chains. PSF-CMV-MUKAPPA – mouse kappa light chain antibody plasmid encodes the constant region from the mouse kappa light chain antibody gene and is designed to allow the fusion of variable antibody gene segments seamlessly to create light chain cassettes. This is possible because there is a BseRI restriction enzyme site upstream of the kappa coding sequence that when cleaved produces an overhang from the first two nucleotides of the first codon of the constant region. By designing the variable regions of antibodies to have a BseRI site at the end in an opposing direction it is possible to seamlessly fuse the variable and constant domains together.

Promoter Expression Level: This plasmid contains the mammalian CMV promoter to drive gene expression. We have tested all of our mammalian promoters in a range of cell types and CMV is consistently the strongest in those we have studied. However there are many reports of the CMV promoter demonstrating silencing by methylation in long-term culture.

應用

PSF-CMV-MUKAPPA – mouse kappa light chain antibody plasmid encodes a constant region an antibody in the main multiple cloning site positioned so that it can be cleaved to produce an overhang that allows seamless fusion with a variable region from any antibody. This allows you to create full length antibody genes with no cloning scars.

To enable this immediately upstream of the constant region coding sequence there is a BseRI restriction site. This is a type-IIS restriction enzyme that binds in one position (CAGCAG) and then cleaves a specific number of nucleotides away from the binding site regardless of the sequence at the cleavage point. We use this site in all of our antibody expression cassettes in the same position. In this plasmid cutting with BseRI will result in an overhang consisting of the first two nucleotides of the first codon of the constant region. This means that any variable region with the same overhang at its 3 prime end can be ligated into this plasmid when used in conjunction with any 5 prime site (NotI-NcoI). To add this overhang the variable region must be PCR amplified to contain any of the following sites at its 3 prime end: BseRI BsgI BtsI or BsrDI. By using this system it allows antibody variable regions to PCR amplified and fused to any of our constant region plasmids without having to re-synthesise the entire antibody expression cassette each time.

序列

To view sequence information for this product, please visit the product page

分析報告

To view the Certificate of Analysis for this product, please visit www.oxgene.com

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儲存類別代碼

12 - Non Combustible Liquids

閃點(°F)

Not applicable

閃點(°C)

Not applicable


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