多系统表达载体pTriEX系列pTriEx Selective Marker Vectors-BioVector NTCC Inc.

价　　格：¥4920
货　　号：多系统表达载体pTriEX
产　　地：北京

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pTriEx™-1.1 Neo DNA

Cat. No. 70927-3

pTriEx Selective Marker Vectors

Efficient selection of stable mammalian cell lines expressing genes of interest Novagen's new selective marker pTriEx™ expression vectors are designed to efficiently select stable mammalian cells expressing target genes. This is achieved by translation of both the gene of interest and a downstream selective marker gene from a single messenger RNA, which ensures that all drug-resistant cells also produce the target protein. Translation of the selective marker gene is controlled by the EMCV-derived Cap-Independent Translation Enhancer (CITE, or IRES, internal ribosome entry site; 1, 2). After selection with neomycin or hygromycin, virtually all surviving cells exhibit stable high level expression of target gene, eliminating the need to screen a large number of surviving colonies.

1. Jang, S.K., Krausslich, H., Nicklin, M.J.H., Duke, G.M., Palmenberg, A. C. and Wimmer, E. (1998) J. Virol. 62, 2636–2643.
2. Jackson, R.J., Howell, M.T. and Kaminski,A. (1990) Trends Biochem. Sci. 15, 477–483.

The b-galactosidase gene from E. coli was cloned into pTriEx-1.1 Neo. The resulting plasmid was transfected into BHK cells by using GeneJuice™ Transfection Reagent. Forty-eight hours post-transfection, neomycin (500 µg/ml) was added to the culture medium. After 7 days of further culture, surviving cells were trypsinized, seeded into a new T-75 flask and grown in the presence of 1000 µg/ml neomycin for an additional 2 weeks. To assess expression of the b-galactosidase target gene, cells were stained with X-gal substrate using a standard protocol. The panels are photographs of the same area in low and high magnification, respectively. Virtually all of the cells in the polyclonal population exhibited high levels of b-gal expression.

Product Description

The pTriEx™-1.1 Neo vector¹ is designed to allow rapid characterization of target genes in multiple expression systems, and to allow rapid selection of stable transfected mammalian cells expressing high levels of target protein. With this vector a single recombinant plasmid can be used to test expression in E. coli, insect and vertebrate cells. Expression in vertebrate cells is mediated by a hybrid promoter composed of the CMV immediate early enhancer fused to the chicken b-actin promoter. The drug selection marker is expressed under the control of the EMC virus Cap-Independent Translation Enhancer (CITE) sequence (or IRES), allowing rapid selection of transfected mammalian cells using the drug G418 or neomycin sulfate. For expression in insect cells, pTriEx-1.1 Neo contains flanking baculovirus sequences to permit the generation of recombinant baculoviruses using the BacVector^® System. In baculovirus-infected insect cells, expression is driven by the very late p10 promoter. Expression in E. coli is regulated by the tightly controlled T7lac promoter. Expression can be induced in hosts such as NovaBlue by infecting with lCE6, a phage that constitutively expresses T7 RNA polymerase from the lp_L and lp_I promoters. Alternatively, pTriEx recombinant plasmids can be transferred into a (DE3)pLacI host that allows induction with IPTG.

•pTriEx-1.1 Neo Sequence
•pTriEx-1.1 Hygro/Neo pTriEx-2 Hygro/Neo Diagrams
•TB293 pTriEx-1.1 Neo Vector Map
•TriEx System Overview
•TB250 TriEx System Manual
•Features of pTriEx Bicistronic Vectors