Prokaryotic Expression and Antiserum Preparation of Papaya Ringspot Virus-Watermelon Strain Coat Protein
Huang Xiande
Wang Yu
Tian Yanping
Gu Qinsheng
Li Xiangdong
Abstract：The coat protein (CP) gene of papaya ringspot virus-watermelon strain (PRSV-W) was amplified by RT-PCR from infected zucchini leaf samples and cloned into the prokaryotic expression vector pEHISTEV to produce recombinant plasmid pEHISTEV-PRSV-W-CP.The recombinant plasmid was transformed into Escherichia coli strain Rosetta and expressed a fusion protein with molecular weight of 37 kD after induction with IPTG.The fusion protein was cut from the gel,emulsified and used to immunize the New Zealand rabbits to produce polyclonal antiserum against the CP of PRSV-W.The titer of the resultant antiserum was 1∶8192 in ELISA.In Western blot analysis,the antiserum showed specific positive reaction only with the zucchini plants infected with PRSV,but not with healthy zucchini plants or those infected with Watermelonmosaic virus,or Nicotiana tabacum plants infected with Potato virus Y.This research laid foundations for the detection of PRSV and functional studies of PRSV CP.
Keywords：Papaya ringspot virus-watermelon strainCoat proteinProkaryotic expressionAntiserum preparationDetection
Publication Date：2017-01-01
Online Publishing Date：2026-05-22(First online date of this platform, not the publication date of the document)
Pages：4( 86-89 )