US 8,568,978 B1 · Use of mammalian expression vector including T7 promoter and N-terminal HA tag for overexpression of human genes in E. coli

Abstract

The present invention relates to a technique for the T7 promoter-driven expression of a foreign protein, preferably, a human protein in E. coli having no T7 RNA polymerase. More particularly, the present invention relates to use of a vector including a T7 promoter and an N-terminal HA tag sequence for overexpression of a foreign protein in E. coli having no T7 RNA polymerase, E. coli that includes the vector and is able to overexpress the foreign protein in the absence of T7 RNA polymerase, a method for expressing or producing the foreign protein using the E. coli, and a method for analyzing functions of the foreign protein. Further, the present invention relates to a simple, rapid method for evaluating the expression and cytotoxicity of a human protein in E. coli by using the vector.

Claims7

Dependent claims

A method for screening a cytotoxic protein, comprising the steps of:

introducing E. coli that does not contain a T7 RNA polymerase with a vector that sequentially includes a T7 promoter, a nucleotide encoding a HA tag, and a gene encoding a foreign protein in the 5′ to 3′ direction;
comparing growth of E. coli harboring the vector to that of a control strain introduced with no vector;
and
determining the foreign protein expressed by the vector as a cytotoxic protein when the growth of E. coli harboring the vector is lower than that of the control strain introduced with no vector.

1 more element

Dependencies are inferred from the claim text.

Facts

Application

13/898,700Filed 21 May 2013 · division

Priority

KR 10-2012-008271827 Jul 2012

Granted

29 Oct 2013

Term ends

Not estimatedThe source records don’t support an estimate.

Classification7 codes

C07K 14/52
From animals; from humans: cytokines; Lymphokines; Interferons
C07K 14/4747
Not used: apoptosis related proteins
C07K 14/7151
For cytokines; for lymphokines; for interferons: for tumor necrosis factor [TNF], for lymphotoxin [LT]
C12P 21/02
Preparation of peptides or proteins: having a known sequence of two or more amino acids, e.g. glutathione

Use of mammalian expression vector including T7 promoter and N-terminal HA tag for overexpression of human genes in E. coli

Life of this patent

Use of mammalian expression vector including T7 promoter and N-terminal HA tag for overexpression of human genes in E. coli

Life of this patent

Abstract

Claims7