人源轮状病毒vp7基因的克隆与转基因植物研究

目的：克隆人源轮状病毒外壳蛋白vp7基因，以制备转基因植物疫苗。方法：用RT-PCR方法制备vp7基因，以植物高效表达质粒PBI121为载体，构建重组DNA质粒。重组体用载体上的通用引物为测序引物，鉴定克隆的正确性。再将鉴定过的重组质粒用农杆菌介导法转染马铃薯外植体，分别用PCR、Western blot法鉴定阳性转化植株中vp7基因的表达。结果：成功地将vp7转入马铃薯植株中，并且在转化植株中检测出了vp7的表达。结论：成功地构建了重组PBI121/hvp7质粒，获得表达外源基因vp7的转基因马铃薯。

Cloning and expressing of human rotavirus vp7 in potatoes

ObjectiveTo clone human rotavirus vp7 gene for prepari ng oral transgenic plant vaccine.MethodsRecombinant DNA was cons tructed by ligating vp7 gene with the vector PBI121 which could be highly expre ssed in plants. The recombinants were verified by sequencing them with the commo n primers on the vector as sequencing primers. Then the correct recombinants wer e transformed into potato exp lants induced by Agrobacterium tumefaciens LB A4404. The expression of vp7 in transgenic potatoes were examined by PCR and Western b lot. Results The vp7 gene has been transformed into potato explants and it expressed in transgenic potatoes permanently. Conclusion We have successfully obtained the transgenic potatoes which expressed the PBI/hvp7 recombinants.