Mutant D178N prion protein converts spontaneously in RT-QuIC assay
10.3760/cma.j.issn.1003-9279.2012.05.015
- VernacularTitle:朊蛋白突变体D178N在RT-QuIC实验中的转化研究
- Author:
Chen GAO
1
;
Ke REN
;
Long-Zhu LI
;
Hui-Ying JIANG
;
Cao CHEN
;
Jin ZHANG
;
Jun HAN
;
Xiao-Ping DONG
Author Information
1. 中国疾病预防控制中心病毒病预防控制所传染病预防控制国家重点实验室
- Keywords:
Prions;
Creutzfeldtfeldt-Jacob Syndrome;
Nucleic acid amplification techniques
- From:
Chinese Journal of Experimental and Clinical Virology
2012;26(5):370-373
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the conversion of mutant D178N prion protein in RT-QuIC assay.Methods The D178N mutant prion PRNP was generated by the method of single site mutation.The mutant PRNP gene was inserted into plasmids of pET24.The full and N-truncated recombinant human prion proteins were expressed and purified.The fibril formations of these proteins were real-time monitored by the method of RT-QuIC.The ability to resist proteinase K (PK) of these fibrils was analyzed.Results We succeed to construct human PrP-D178N plamids.The N-truncated human prion protein with D178N (PrP90-231-D178N) can convert spontaneously in RT-QuIC,while full length of human prion D178N protein(PrP23-231-D178N) fails to convert spontaneously.The spontaneously generated fibril has been domenstrated it is partily PK-resistant.Conclusion The N-terminal of prion protein (23-90) plays an important role for the D178N mutant protein spontaneously conversion,which provide the clues for study the pathogenesis of genetic CJD.
