title: Incidence of Lettuce mosaic virus in lettuce and its detection by polyclonal antibodies produced against recombinant coat protein expressed in Escherichia coli.
abstract: Lettuce mosaic virus (LMV), a member of the genus Potyvirus of family Potyviridae, causes mosaic disease in lettuce has recently been identified in India. The virus is seed borne and secondary infection occurs through aphids. To ensure virus freedom in seeds it is important to develop diagnostic tools, for serological methods the production of polyclonal antibodies is a prerequisite. The coat protein (CP) gene of LMV was amplified, cloned and expressed using pET-28a vector in Escherichia coli BL21DE3 competent cells. The LMV CP was expressed as a fusion protein containing a fragment of the E. coli His tag. The LMV CP/His protein reacted positively with a commercial antiserum against LMV in an immunoblot assay. Polyclonal antibodies purified from serum of rabbits immunized with the fusion protein gave positive results when LMV infected lettuce (Lactuca sativa) was tested at 1:1000 dilution in PTA-ELISA. These were used for specific detection of LMV in screening lettuce accessions. The efficacy of the raised polyclonal antiserum was high and it can be utilized in quarantine and clean seed production.
authors: Advanced Centre for Plant Virology, Division of Plant Pathology, Indian Agricultural Research Institute (IARI), New Delhi 110012, India. Electronic address: prachisharma15@gmail.com.
authors: Prachi
authors: P
authors: Sharma
authors: School of Biotechnology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu (SKUAST-J), Jammu and Kashmir 180009, India.
authors: Susheel
authors: S
authors: Sharma
authors: Advanced Centre for Plant Virology, Division of Plant Pathology, Indian Agricultural Research Institute (IARI), New Delhi 110012, India.
authors: Jasvir
authors: J
authors: Singh
authors: Division of Vegetable Sciences, IARI, New Delhi 110012, India.
authors: Swati
authors: S
authors: Saha
authors: Advanced Centre for Plant Virology, Division of Plant Pathology, Indian Agricultural Research Institute (IARI), New Delhi 110012, India.
authors: V K
authors: VK
authors: Baranwal
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D018545:Lactuca / Q000821:virology*; D017800:Potyvirus / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology*; D012698:Serologic Tests
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins
keywords: Lactuca sativa; Mosaic; PTA-ELISA; Polyclonal antibodies; Potyviridae; Western blot