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Title 

An efficient expression vector for extracellular secretion in mammalian cells

Authors 

Young Choon LeeChul Ho KimShuichi Tsuji

Publisher 

Springer Verlag (Germany)

Issue Date 

1996

Citation 

Molecules and Cells, vol. 6, no. 5, pp. 552-556

Abstract 

An expression-secretion vector for mammalian cells, pcDSA, which expresses a cloned gene under the control of the SRa promoter (SV40 promoter/enhancer and HTLV-1 LTR) has been newly constructed. This vector contains fragments encoding the 5′ untranslated leader sequence from AMV RNA4, the signal peptide of mouse IgM and IgG-binding domain of protein A in front of cloning sites. Joining in-frame a cDNA fragment with cloning sites just downstream of the COOH terminus of the IgG-binding domain of protein A enables the cDNA product to be secreted as a protein fused with that domain. This allows an easy isolation of its secreted product by affinity chromatography on IgG-Sepharose. When the genes encoding the catalytic domains of mammalian sialyltransferase (ST3Gal I) were cloned into the vector plasmid and then transfected into COS-7 cells, active ST3Gal I was efficiently secreted into the culture medium. It warf rapidly purified almost to homogeneity by one-step IgG-Sepharose affinity chromatography.

ISSN 

1016-8478

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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