title: Recombination and expression of classical swine fever virus (CSFV) structural protein E2 gene in Chlamydomonas reinhardtii chroloplasts.
abstract: 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.
authors.affiliation: Department of Biotechnology, College of Life Science, Zhejiang University, Hangzhou 310027, China.
authors.forename: Dong-Mei
authors.initials: DM
authors.lastname: He
authors.forename: Kai-Xian
authors.initials: KX
authors.lastname: Qian
authors.forename: Gui-Fang
authors.initials: GF
authors.lastname: Shen
authors.forename: Zhi-Fang
authors.initials: ZF
authors.lastname: Zhang
authors.forename: Yi-Nü
authors.initials: YN
authors.lastname: Li
authors.forename: Zhong-Liang
authors.initials: ZL
authors.lastname: Su
authors.forename: Hong-Bo
authors.initials: HB
authors.lastname: Shao
mesh_terms: 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
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D014759:Viral Envelope Proteins; C087086:glycoprotein E2, classical swine fever virus