High level expression of a functionally active cholera toxin B: rabies glycoprotein fusion protein in tobacco seeds.
A synthetic DNA construct containing cholera toxin B subunit, genetically fused to the surface glycoprotein of rabies virus was expressed in tobacco plants from a seed specific (legumin) promoter. Seed specific expression was monitored by real-time PCR, GM1-ELISA and Western blot analyses. The fusion protein accumulated in tobacco seeds at up to 1.22% of the total seed protein. It was functionally active in binding to the GM1-ganglioside receptors, suggesting its assembly into pentamers in seeds of the transgenic plants. Immunoblot analysis confirmed that the approximately 80.6 kDa monomeric fusion polypeptide was expressed in tobacco seeds and accumulated as an approximately 403 kDa pentamer. Evaluation of its immunoprotective ability against rabies and cholera is to be examined.
Tiwari
Siddharth
S
Plant Molecular Biology and Genetic Engineering Division, National Botanical Research Institute, Council of Scientific and Industrial Research, Rana Pratap Marg, Lucknow 226001, India.
Mishra
Devesh K
DK
Roy
Sribash
S
Singh
Ankit
A
Singh
P K
PK
Tuli
Rakesh
R
D015153:Blotting, Western; D002772:Cholera Toxin / Q000378:metabolism*; D003001:Cloning, Molecular; D018744:DNA, Plant / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D018506:Gene Expression Regulation, Plant; D018745:Genome, Plant / Q000235:genetics; D006023:Glycoproteins / Q000378:metabolism*; D015151:Immunoblotting; D030821:Plants, Genetically Modified; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011820:Rabies virus / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D012639:Seeds / Q000235:genetics*; D014026:Nicotiana / Q000235:genetics*; D019076:Transgenes / Q000235:genetics; D014764:Viral Proteins / Q000378:metabolism*
D018744:DNA, Plant; D006023:Glycoproteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D002772:Cholera Toxin