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Constructing the recombinant of pET28a-TAT-LacZ

Yan Shi

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Abstract

A synthesized double-stranded oligomeric nucleotide encoding 11-amino acid TAT protein transduction domain3 was inserted into pET28a vector after 6 histidind coding sequence,LacZ gene from pcDNA4/Myc-His/LacZ was digested by EcoRⅠand HindⅢ,then cloned into pET28a-TAT.The highly expressed TAT-β-galactosidase was purified by affinity chromatography.TAT-β-Gal fusion protein can across vascular smooth muscle cells in vitro.This result open new possibility for direcy delivery of protein into tissues for therapy.

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What this paper is about

A synthesized double-stranded oligomeric nucleotide encoding 11-amino acid TAT protein transduction domain3 was inserted into pET28a vector after 6 histidind coding sequence,LacZ gene from pcDNA4/Myc-His/LacZ was digested by EcoRⅠand HindⅢ,then cloned into pET28a-TAT.The highly expressed TAT-β-galactosidase was purified by affinity chromatography.TAT-β-Gal fusion protein can across vascular smooth muscle cells in vitro.This result open new possibility for direcy delivery of protein into tissues for therapy.

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Available abstract

A synthesized double-stranded oligomeric nucleotide encoding 11-amino acid TAT protein transduction domain3 was inserted into pET28a vector after 6 histidind coding sequence,LacZ gene from pcDNA4/Myc-His/LacZ was digested by EcoRⅠand HindⅢ,then cloned into pET28a-TAT.The highly expressed TAT-β-galactosidase was purified by affinity chromatography.TAT-β-Gal fusion protein can across vascular smooth muscle cells in vitro.This result open new possibility for direcy delivery of protein into tissues for therapy.

Key concepts: lac operon, Molecular biology, Recombinant DNA, Fusion protein, Affinity chromatography, Gene, Coding region, Transduction (biophysics)

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