2013•Journal of Food Science and BiotechnologyRequires access

Expression and Purification of Cell Penetrating Peptide TAT and Mouse Stem Cell Transcriptional Factor Oct4 Recombinant Protein

Ju Wang

Open publisher page 0 citations

Abstract

In this study,TAT-Oct4 target gene was inserted into pET28a expression vector to construct pET28a-TAT-Oct4 recombinant expression vector. TAT-Oct4 fusion protein was mainly expressed with insoluble inclusion bodies in E.coli. The purity of expression product was more than90% after purification. And the average refolding field is 8.8% using urea gradient dialysis.Immunocytochemistry analysis showed that TAT-Oct4 recombinant protein could penetrate nearly100% cells membrane and was mainly concentrated in nucleus. The method of generating active TAT-Oct4 recombinant protein was established in this article. It provided not only effective study materials for cell reprogramming but also practical method for other reprogramming factors design.

About this research paper

What this paper is about

In this study,TAT-Oct4 target gene was inserted into pET28a expression vector to construct pET28a-TAT-Oct4 recombinant expression vector. TAT-Oct4 fusion protein was mainly expressed with insoluble inclusion bodies in E.coli. The purity of expression product was more than90% after purification. And the average refolding field is 8.8% using urea gradient dialysis.Immunocytochemistry analysis showed that TAT-Oct4 recombinant protein could penetrate nearly100% cells membrane and was mainly concentrated in nucleus. The method of generating active TAT-Oct4 recombinant protein was established in this article. It provided not only effective study materials for cell reprogramming but also practical method for other reprogramming factors design.

Why it matters

A significance statement is not available in the OpenAlex record.

Key contribution

A contribution statement is not available in the OpenAlex record.

Method / approach

Method details are not available in the OpenAlex metadata.

Main findings

Findings are not separately available in the OpenAlex metadata.

Limitations

Limitations are not available in the OpenAlex metadata.

Applications

Application details are not available in the OpenAlex metadata.

Available abstract

In this study,TAT-Oct4 target gene was inserted into pET28a expression vector to construct pET28a-TAT-Oct4 recombinant expression vector. TAT-Oct4 fusion protein was mainly expressed with insoluble inclusion bodies in E.coli. The purity of expression product was more than90% after purification. And the average refolding field is 8.8% using urea gradient dialysis.Immunocytochemistry analysis showed that TAT-Oct4 recombinant protein could penetrate nearly100% cells membrane and was mainly concentrated in nucleus. The method of generating active TAT-Oct4 recombinant protein was established in this article. It provided not only effective study materials for cell reprogramming but also practical method for other reprogramming factors design.

Key concepts: Recombinant DNA, Molecular biology, Fusion protein, Myc-tag, Reprogramming, FLAG-tag, Expression vector, Biology

Related papers

Back to paper searchBrowse research topicsOriginal source
Expression and Purification of Cell Penetrating Peptide TAT and Mouse Stem Cell Transcriptional Factor Oct4 Recombinant Protein — Research Paper | ScholarLens