Constructing And Expressing As Well As Anti-Serum Preparation Of The CP And GFP Fursion Gene In Escherichia Coli

Barley yellow dwarf viruses(BYDVs),transmitted naturally by at least 25 aphid species in a highly specific,circulative,non—propagative manner,are a kind of +ssRNA viruses.Barley yellow dwarf viruses(BYDVs) caused severely yellowing and dwarfing on wheat,barley and oat.BYDVS-PAV can be transmitted efficiently by the Rhopalosiphum padi Linnaeus and Sitobion(Macrosiphum) avenae.According to published nucleotide sequences ,CP gene of barley yellow dwarf virus PAV (BYDV-PAV) was synthesized by reverse transcription polymerase chain reaction (RT-PCR).This experiment put the reporter gene GFP to the downstream of whole gene sequence(CP)of barley yellow dwarf virus PAV strain's coat protein, after double enzyme digestion,PCR identification and sequencing,we has uccessfully constructed the green fluorescence protein expression system of coat protein of barley yellow dwarf virus PAV strain and expressed in E. Coli BL21(DE3)Plys S. Its expression product was confirmed by sds-page,western bolt analysis and fluorescence microscope.The fusion gene CP-GFP expressed in the range of 15-22℃, but cannot expressed from 23℃to 37℃.These results provide a base for BYDV-PAV in aphid's vivo movement further.Applying the target protein(CP-GFP- Pro) as antigen to immune rabbit, the antiserum was obtained.Potency that is 1:512 was confirmed by agar double diffusion technique.