The Expression Of Human Rotavirus In Carrot

Carrot was applied in the experiment as receptor of bioreactor to produce RV protein VP6, VP7. And transgenic carrots in different generations were screened with the methods of PCR,RT-PCR,Western Blot,ELISA. The transgenic carrots expressing RV protein were applied to immune mice.1 The PCR analysis showed that VP6,VP7,VP7-U gene had integrated to carrot genome.2 The result of RT-PCR demonstrated that only some transgenic carrots had reverse transcriptase procedure.3 Western Blot and ELISA showed that VP7 protein had been expressed effectively in transgenic carrots and the product had a certain immunogenicity.4 The root protein of transgenic carrot with VP7 gene was applied to immune mice through injection together with freund's adjuvant and the dosage of antigen was about 1μg. The immunization period was two weeks and mice blood was got to extract serum after three times immunization. The ELISA analysis for serum showed that the content of IgG in the mice injecting positive carrot root protein was higher than that of the mice injecting negative carrot root protein or non. The root protein of transgenic carrot with VP7-U gene was extracted to immune mice through filling stomach after analysises of PCR, RT-PCR,Western Blot,ELISA,and the antigen dosage was 20μg to 30μg. The immunization period was one week and mice blood was got to extract serum after four times immunization for ELISA analysis. The ELISA result showed that the content of IgG in the mice injecting positive carrot root protein was higher than that of the mice injecting negative carrot root protein or non.5 Comparing with the carrots transformed VP7 gene, the antigen content in carrots transformed VP7-U gene was higher.