Foot-and-mouth disease virus VP1 protein fused with cholera toxin B subunit expressed in Chlamydomonas reinhardtii chloroplast.
A Chlamydomonas reinhardtii chloroplast expression vector, pACTBVP1, containing the fusion of the foot and mouth disease virus (FMDV) VP1 gene and the cholera toxin B subunit (CTB) gene was constructed and transfered to the chloroplast genome of C. reinhardtii by the biolistic method. The transformants were identified by PCR, Southern blot, Western blot and ELISA assays after selection on resistant medium and incubation in the dark. The CTBVP1 fusion protein was expressed in C. reinhardtii chloroplast and accounted for up to 3% of the total soluble protein. The fusion protein also retained both GM1-ganglioside binding affinity and antigenicity of the FMDV VP1 and CTB proteins. These experimental results support the possibility of using transgenic chloroplasts of green alga as a mucosal vaccine source.
Sun
Meng
M
Department of Biotechnology, College of Life Science, Zhejiang University, Hangzhou 310027, PR China. sunmeng1976@sohu.com
Qian
Kaixian
K
Su
Ning
N
Chang
Huiyun
H
Liu
Jixing
J
Shen
Guifang
G
Chen
Guifang
G
D000818:Animals; D019470:Biolistics / Q000379:methods; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D016825:Chlamydomonas reinhardtii / Q000235:genetics* / Q000378:metabolism*; D002736:Chloroplasts / Q000235:genetics / Q000378:metabolism*; D002772:Cholera Toxin / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005786:Gene Expression Regulation; D030781:Organisms, Genetically Modified / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D014170:Transformation, Genetic; D014614:Vaccines, Synthetic / Q000096:biosynthesis / Q000235:genetics
D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C442501:VP1 protein, Foot-and-mouth disease virus; D014614:Vaccines, Synthetic; D002772:Cholera Toxin