The Codon-optimized Rotavirus Vp6 Protein In Tobacco Plants Highly Expressed

Rotavirus (RV) is the most important and common cause of severe diarrhea in infants, children and infancy animal worldwide. In developing countries, rotavirus may cause an estimated 18 million cases of moderately severe and severe diarrhea and over 870 000 deaths in the young each year. In developed countries, rotavirus may resulted in substantial economical damage. For example, medical expenditure associated with rotavirus may reach 260 000 000＄each year, in the United States. Because of significant morbidity and mortality associated with rotavirus diarrhea, there is an urgent need to develop rotavirus vaccines targeted for use in early infancy. Currently.Plant reactor system is cheap and safe biosystem. With the development of the Molecular biology, and especially the fast step of plant genetic engineering, people can rebuild the plant in their own opinions, and can produce many kinds of product needed in production and life cheaply and efficiently. The concept of plant reactor, and especially the concept of producing officinal protein and enzyme with plant and using plant directly as the oral vaccine, is gradually recognized and accepted by human being.In this experiment, we chose tobacco to produce VP6 protein. The codon optimized VP6 gene was insert into plasmid pBI121 int, and the plant transformation vector (pBI121-V6(U)) was constructed to express the VP6 protein. By using the leaf-disc co-cultivated method , VP6 gene of rotavirus was transferred into tobacco cell respectively. Transformed shoots were selected on solidified medium Containing Kanamycin. Integration of these genes into the genome of tobacco plants was confirmed by PCR. And these results indicated that the VP6 was integrated into the tobacco genomic DNA. The western blot result showed that the protein was expressed efficiently. This research provides a new method for new rotavirus vaccine, which presents theorical significance and applicative prospect.