Preparation of Protein Transduction Domain Modified Mice IFN-?
- VernacularTitle:蛋白转导结构域修饰小鼠干扰素-?的研制
- Author:
Daqiang HU
- Publication Type:Journal Article
- Keywords:
Protein transduction domain;
IFN-?;
Protein purification;
ELISA
- From:
China Pharmacy
1991;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To prepare PTD modified mice interferon gamma(PTD-mIFN-?) in order to prepare for its function research.METHODS:The code of PTD-mIFN-? fragment was amplified by PCR for 3 times,using pUC19/mIFN-? as template.The PCR product was cloned into pUC19 plasmid and then constructed into expression plasmid vector pQE80L after sequencing.After double digestion with BamH Ⅰ and Hind Ⅲ,the recombinant vector was identified in 1.2% agarose gel.The target protein and PTD-mIFN-? was checked by SDS-PAGE,Western blot method and ELISA assay,respectively.Recombinant protein was purified by nikel ion-triglycollamic acid(Ni2+-NTA) Agarose affinity chromatogram.RESULTS:The cDNA fragment encoding PTD-mIFN-? amplified by PCR was obtained.The high-level expression of recombinant protein was noted in SDS-PAGE and Western blot assay.The target protein was obtained by ELISA assay.CONCLUSION:PTD-mIFN-? was prepared successfully.
