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《Chinese Journal of Microecology》 2017-08
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Expression and purification of Norovirus capsid protein VP1 and preparation of the polyclonal antibodies

ZHANG Qiushi;YUAN Rongliang;JING Shenrong;Medical Faculty,Kunming University of Science and Technology;  
Objective To obtain purified Norovirus capsid protein VP1,and prepare the polyclonal antibodies against VP1.Methods The RNA of Norovirus strain preserved in laboratory was extracted and reverse transcribed.VP1 gene sequence was obtained by PCR amplification and constructed in the prokaryotic expression system of E.coli to induce the expression of recombinant VP1 protein.The recombinant protein was purified by using nickel column affinity chromatography,and analyzed with SDS-PAGE and BSA methods.With the recombinant VP1 protein as the antigen,male SD rats(SPF)were immune to get the antiserum,which was detected for the titer by using ELISA,and for the specificity by using Western blot.Results The recombinant expression vector VP1-pET28 awas successfully constructed,and stably expressed in E.coli BL21(DE3).The average titer of the polyclonal antibody was 1∶200 000 by ELISA.Western blot confirmed high prokaryotic and eukaryotic specificity of the antibodies.Conclusion With the success of the prokaryotic expression system of Norovirus capsid protein VP1,the study provided a basis for further study on the diagnosis of Norovirus and development of the vaccine.
【CateGory Index】: R392
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