Recombination and expression of classical swine fever virus (CSFV) structural protein E2 gene in Chlamydomonas reinhardtii chroloplasts.
The expression of classical swine fever virus (CSFV) structural protein E2 in different vectors, which has been shown to carry critical epitopes, has been established. Here, we reported a Chlamydomonas reinhardtii chloroplast expression vector, P64E2, containing classical swine fever virus structural protein E2 gene, which was constructed and transferred to C. reinhardtii by biolistic bombardment method. The transformants were identified by PCR, Southern blotting, Western blotting after selecting on resistant media. ELISA quantification assay showed that the expressed E2 protein accumulated up to 1.5-2% of the total soluble protein. The results of the study on the immunological activity indicated that the protein E2 expressed in C. reinhardtii chloroplasts could elicit animal bodies to produce antibodies against protein E2.
He
Dong-Mei
DM
Department of Biotechnology, College of Life Science, Zhejiang University, Hangzhou 310027, China.
Qian
Kai-Xian
KX
Shen
Gui-Fang
GF
Zhang
Zhi-Fang
ZF
Li
Yi-Nü
YN
Su
Zhong-Liang
ZL
Shao
Hong-Bo
HB
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D015153:Blotting, Western; D016825:Chlamydomonas reinhardtii / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D018014:Gene Transfer Techniques; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic / Q000235:genetics* / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D014759:Viral Envelope Proteins; C087086:glycoprotein E2, classical swine fever virus