NR ANNM
AU Zhao,X.; Dong,X.; Hung,T.
TI [Expression of human prp gene in prokaryotic cells using GST fusion protein expression system]
QU Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese Journal of Experimental and Clinical Virology 1999 Jun 30; 13(2): 124-7
PT journal article
AB OBJECTIVE: To study the biological features of cell-surface protein PrPc, which is thought to be involved in the prion-associated diseases after converting to a proteinase-resistant isoform PrPsc posttranslationally, and to establish an effective immunologic diagnostic method using PrPc as antigen. METHODS: Amplifying and cloning the human prp gene from lymphocytes of two normal Chinese, after confirmed by DNA sequence analysis, the genes were separately subcloned into a GST-fusion expression plasmid. RESULTS: Sequence analysis showed that one contatined a point mutation that induced the 65th amino acid "Trp" inverting to a stop codon "TAG", whereas the other had the same sequence as the published standard prp gene. Both the standard and the mutated prp genes were separately subcloned into a GST fusion protein expression vector and expressed in the prokaryotic cells effectively. Western blot assay revealed that both of them expressed GST-PrP fusion proteins and could be recognized by PrP specific monoclonal antibody. CONCLUSION: It suggests that human PrP protein can be expressed in the GST fusion protein expression system and the expressed proteins hold good immune-reactivity.
AD Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052.
SP chinesisch
PO China