[Transgenic potato containing immunogenic gene of avian coronavirus and its immunogenicity in mice].
To check the feasibility of expression of the immunogenic gene of avian coronavirus infectious bronchitis virus (IBV) in plants, the transformation of S1 gene of IBV into potato and the immunogenicity of its expression product was studied. The S1 gene of IBV-ZJ971 strain was inserted into plasmid pBI121 under the control of 35 S promoter. Agrobacterium fumefaciens EHA105 with the recombinant vector pBI121 was obtained by tri-parental mating method. So, an efficient potato transformation system mediated by Agrobacterium fumefaciens was established. The rates of calli and shoots differentiation were 100%, and more than 95% respectively, for transgenic potato with S1 gene of IBV. PCR and Southern blot analyses showed that IBV S1 gene was integrated into genomic DNA of the potato plant and most transgenic plants had two copies of S1 gene of IBV. In our experiments, 47 transgenic plantlets have been obtained. Northern blot and ELISA analyses indicated that most transgenic plants could normally transcribe and translate S1 gene of IBV, though the levels of transcription and translation were different in various transgenic plants. Immunity assay with BALB/C mice showed that expression products of transgenic potato with S1 gene of IBV were immunogenic, and ELISA antibody titer reached 1:20 to 1:40 and 1:80 to 1:160 with doses of 0.5 g and 1 g, respectively. Virus neutralization (VN) antibodies were detected by tracheal organ cultures, and the results showed that VN titers reached respectively 1:160 to 1:320 and 1:320 to 1:2048 with doses of 0.5 g and 1 g.
Wu
Jian-Xiang
JX
Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou 310029, China.
Zhou
Ji-Yong
JY
Zhou
Xue-Ping
XP
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D015152:Blotting, Northern; D015139:Blotting, Southern; D046508:Culture Techniques; D018744:DNA, Plant / Q000235:genetics; D015870:Gene Expression; D001351:Infectious bronchitis virus / Q000235:genetics* / Q000276:immunology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D030821:Plants, Genetically Modified; D018749:RNA, Plant / Q000235:genetics; D011198:Solanum tuberosum / Q000235:genetics*; D064370:Spike Glycoprotein, Coronavirus; D014170:Transformation, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology
D000914:Antibodies, Viral; D018744:DNA, Plant; D008562:Membrane Glycoproteins; D018749:RNA, Plant; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins