HV single-chain antibody - protamine fragment fusion protein 1A8 ScFv - Cκ - P Construction and expression of.docVIP

 PAGE \* MERGEFORMAT 13 HV single-chain antibody / protamine fragment fusion protein 1A8 ScFv / Cκ / P Construction and expression of Author: Chen Rui, Yang, Fang-Lin Zhang, Qin Wei Wei, Wang Tao, Zhang Rui, Meng Yanling, Hu Gang, XU Zhi-kai, YANG Gang [Abstract] Objective: To construct Hantaan virus (Hantavirus, HV) single-chain antibody / protamine fragment fusion protein 1A8 ScFv / Cκ / P prokaryotic expression vector, protein was purified after expression and conduct 1A8 ScFv / Cκ / P fusion identification of protein functional activity. Methods: The PCR method, amplified single-chain antibody gene 1A8 ScFv and 1A8 ScFv / Cκ , the protamine fragment coding sequences into the primers designed to obtain HV single-chain antibody / protamine The fusion protein gene fragment 1A8 ScFv / Cκ / P. will be the recombinant gene was cloned into the prokaryotic expression vector pET32a, and expressed in E. coli BL21. expression product by SDS  PAGE and Western Blot identification, with the layer of Ni  NTA chelate Analysis of the purified medium. using ELISA method of analysis of 1A8 ScFv / Cκ / P fusion protein with the HV antigen binding activity, gel mobility shift experiments detected 1A8 ScFv / Cκ / P fusion protein with the plasmid DNA-binding activity. Results: The successful construction the HV 1A8 scFv / Cκ / P fusion protein gene expression vector in E. coli expression of soluble achieved. through functional activity assay, purified 1A8 scFv / Cκ / P fusion protein with the HV while maintaining the antigen binding capacity, while Also has the nucleic acid binding activity. Conclusion: The Construction and expression of the HV 1A8 scFv / Cκ / P fusion protein with the HV antigen binding activity and DNA binding activity. [Keywords:] Hantaan virus; single-chain antibody; protamine; fusion protein; targeted delivery 0 Introduction Selective transfer of genes into the body, various types of cells and organs can be of human diseases, particularly