Polyclonal antibodies against the recombinantly expressed coat protein of the Citrus psorosis virus
Joint Authors
Salamah, Muhammad
Salim, Rida
Uthman, Jamal E. H.
Arif, Ibrahim Abd al-Wahid
Source
Saudi Journal of Biological Sciences
Issue
Vol. 25, Issue 4 (31 May. 2018), pp.733-738, 6 p.
Publisher
Publication Date
2018-05-31
Country of Publication
Saudi Arabia
No. of Pages
6
Main Subjects
Abstract EN
Psorosis is a damaging disease of citrus that is widespread in many parts of the world.
Citrus psorosis virus (CPsV), the type species of the genus Ophiovirus, is the putative causal agent of psorosis.
Detection of CPsV by laboratory methods, serology in particular is a primary requirement for largescale surveys but their production has been impaired by the difficulty of obtaining sufficient clean antigen for immunization.
Specific PAbs against coat protein were produced in E.
coli using recombinant DNA approach.
The full length CP gene fragment was amplified by RT-PCR using total RNA extracted from CPsV infected citrus leaves and CP specific primers.
The obtained product (1320bp) was cloned, sequenced and sub-cloned into pET-30(+) expression vector.
Expression was induced and screened in different bacterial clones by the presence of the expressed protein (48kDa) and optimized in one clone.
Expressed CP was purified using batch chromatography under denaturing conditions.
Specificity of expressed protein was demonstrated by ELISA before used as antigen for raising PAbs in mice.
Specificity of the raised PAbs to CPsV was verified by ELISA and western blotting.
The raised PAbs were showed highly effectiveness in screening by ELISA comparing with the commercial antibodies purchased from Agritest, Valanzano, Italy.
The expression of CPsV CP gene in E.
coli, production of PAbs using recombinant protein as an antigen, the suitability of these antibodies for use in immunodiagnostics against the CPsV Egyptian isolate have been accomplished in this work.
American Psychological Association (APA)
Salim, Rida& Arif, Ibrahim Abd al-Wahid& Salamah, Muhammad& Uthman, Jamal E. H.. 2018. Polyclonal antibodies against the recombinantly expressed coat protein of the Citrus psorosis virus. Saudi Journal of Biological Sciences،Vol. 25, no. 4, pp.733-738.
https://search.emarefa.net/detail/BIM-838799
Modern Language Association (MLA)
Salim, Rida…[et al.]. Polyclonal antibodies against the recombinantly expressed coat protein of the Citrus psorosis virus. Saudi Journal of Biological Sciences Vol. 25, no. 4 (May. 2018), pp.733-738.
https://search.emarefa.net/detail/BIM-838799
American Medical Association (AMA)
Salim, Rida& Arif, Ibrahim Abd al-Wahid& Salamah, Muhammad& Uthman, Jamal E. H.. Polyclonal antibodies against the recombinantly expressed coat protein of the Citrus psorosis virus. Saudi Journal of Biological Sciences. 2018. Vol. 25, no. 4, pp.733-738.
https://search.emarefa.net/detail/BIM-838799
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references : p. 737-338
Record ID
BIM-838799