Animal The Rotavirus Housing Antigen Protein Vp4 Gene Into The Ramie Research

Rotavirus is one of the main pathogen that caused the Virus diarrhea of young animals. The disease of infected animal caused serious economy destruction. Now vaccine precaution is the optical way to control the diarrhea, but the effect of the dead vaccine was not good and the live vaccine would cause the diffusion and environment contaminate. Oral vaccines produced by transgenic plants would change the traditional means of production and inoculation of vaccines, and the cost of vaccine production would be reduced greatly. Consequently it is very significant to develop transgenic plant vaccines.The regeneration system of Boechtneria nivea L. Guad was created in this research. The medium RI of induction calla, R2 medium of induction shoots and RS medium of induction roots was selected by experiment, which built a base for plant transformed.In this research, the VP4 gene was modified by PCR amplification. The kozak sequence was placed before the start codon of VP4, which was advantageous for the gene expression. At the same time, VP4 gene was mutated in a certain point by PCR. The plant expression vector: pBI121 was constructed and then transformed to Agrobacteriutn tumefaciens EHA105 directly. A.tumefaciens EHA105 harboring pBI121/VP4 was used to transform the Boechmeria nivea L. Guad according to the leaf disc procedure. Transformed shoots were selected on solidified medium containing Kanamycin. Ten Kanamycin resistant transformants were obtained by direct or induction calla. These transformants were checked by PCR, PCR-Southern blot and Southern blot , confirmed that two positive transformants were integrated into the genome of Boechmeria nivea L Guad.