Detection for CMV,LSV,LMoV Infected Lily with DNA Microarry Techniques
Abstract: Coat protein (CP) gene of Tobacco ringspot virus (TRSV) Shandong isolate 1 was cloned by reverse transcription-polymerase chain reaction (RT-PCR), and was subcloned into the pET-22b(+) prokaryotic expression vector. The recombinant vector was transformed into E.coli strain BL21. Sequence analysis revealed that the cp gene was 1548 nucleotides in length,encodes a coat protein of 515 amino acids, and shares 90.7%~94.6% nucleotides and amino acid homology with TRSV cp genes registered in GenBank. The target fusion peptide with a molecular weight of 58kDa was expressed under the condition of 23-25℃ and induced by IPTG at a final concentration of 1mmol/L. Rabbit was immunized using the expressed target peptide as antigen, and the antiserum was obtained. The antiserum had a titer of 1/1024 with high specificity to TRSV.