The new method for transfection of macromolecules into cells using protein transductions domain

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Author:
ZHANG Yan-jun(Institute of Hematology,CAMS & PUMC, State Key Laboratory of Experimental Hematology, Tianjin 300020, China)
REN Si-mei()
LU Hong()
LIU Qian()
ZENG Jie()
ZHANG Yi-zhi(Institute of Hematology,CAMS & PUMC, State Key Laboratory of Experimental Hematology, Tianjin 300020, China)
Journal Title:
Journal of Leukemia & Lymphoma
Issue:
Volume 21, Issue 10, 2012
DOI:
10.3760/cma.j.issn.1009-9921.2012.10.002
Key Word:
Protein transductions domain;Transfection

Abstract: Objective To construct a fusion protein to transfect some cell lines that were difficult to be transfected such as neoplastic cells, nerve cells, stem cells. Methods PCR was performed to amplified protein transductions domain(PTD),G4S and streptavidin (Strep).Enzymatic digestion and ligation were used to construct pAYZ-PTD-Strep plasmid. Fusion protein was induced to express by AP5 medium and was isolated by E-tag affinity chromatography. Fusion protein was identified by Western blot. eGFP was trasfected into U937 cells by pAYZ-PTD-Strep. FACS was performed to detect transfection percentage. Results Fusion protein PTD-G4S-Strep was expressed as soluble protein.The concentration of fusion protein was about 0.7 mg/L,and purity was over 90 %. The protein could carry plasmid into a suspended cell line, U937 cells. The transfection-efficiency of protein was higher than monometer PTD.Conclusion The protein PAYZ-PTD-Strep could carry macromolecules into blood tumor cells,and its biological activity may be expected to develop into a highly efficient and reliable transfection method.

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