title: Expression and the antigenicity of recombinant coat proteins of tungro viruses expressed in Escherichia coli.
abstract: Rice tungro disease (RTD) is a recurring disease affecting rice farming especially in the South and Southeast Asia. The disease is commonly diagnosed by visual observation of the symptoms on diseased plants in paddy fields and by polymerase chain reaction (PCR). However, visual observation is unreliable and PCR can be costly. High-throughput as well as relatively cheap detection methods are important for RTD management for screening large number of samples. Due to this, detection by serological assays such as immunoblotting assays and enzyme-linked immunosorbent assay are preferred. However, these serological assays are limited by lack of continuous supply of antibodies as reagents due to the difficulty in preparing sufficient purified virions as antigens. This study aimed to generate and evaluate the reactivity of the recombinant coat proteins of Rice tungro bacilliform virus (RTBV) and Rice tungro spherical virus (RTSV) as alternative antigens to generate antibodies. The genes encoding the coat proteins of both viruses, RTBV (CP), and RTSV (CP1, CP2 and CP3) were cloned and expressed as recombinant fusion proteins in Escherichia coli. All of the recombinant fusion proteins, with the exception of the recombinant fusion protein of the CP2 of RTSV, were reactive against our in-house anti-tungro rabbit serum. In conclusion, our study showed the potential use of the recombinant fusion coat proteins of the tungro viruses as alternative antigens for production of antibodies for diagnostic purposes.
authors.affiliation: Institute of Health and Community Medicine, Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia; Agriculture Research Centre Semongok, Department of Agriculture, Sarawak, Jalan Puncak Borneo, 93720 Kuching, Sarawak, Malaysia.
authors.forename: Siew Fung
authors.initials: SF
authors.lastname: Yee
authors.affiliation: Institute of Health and Community Medicine, Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia.
authors.forename: Chia Huay
authors.initials: CH
authors.lastname: Chu
authors.affiliation: Agriculture Research Centre Tuaran, Department of Agriculture, Sabah, P.O. Box No. 3, 89207 Tuaran, Sabah, Malaysia.
authors.forename: Evenni
authors.initials: E
authors.lastname: Poili
authors.affiliation: Institute of Health and Community Medicine, Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia. Electronic address: shsmag@ihcm.unimas.my.
authors.forename: Magdline Sia Henry
authors.initials: MSH
authors.lastname: Sum
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology*; D001210:Asia, Southeastern; D036022:Capsid Proteins / Q000235:genetics* / Q000276:immunology*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics*; D012275:Oryza / Q000821:virology; D010935:Plant Diseases / Q000821:virology; D012367:RNA, Viral / Q000235:genetics; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology; D019193:Waikavirus / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins
keywords: Antibody; Recombinant fusion protein; Serological assays; Tungro viruses