Sequence Analysis Of Genome Sequence,Ployantibody Against CPm Protein And Its Application Of Lettuce Chlorotic Virus

Lettuce chlorosis virus(LCV),belongs to genus Crinivirus,is newly contagious virus threatening Solanaceae crops in China.LCV possesses similar characteristics of genome structure,transmission with Tomato chlorosis virus(ToCV),which severely epidemic in Solanaceae crops in China.So it is urgently needed to launch the studies including virus distribution,viral pathogenecity to uncover LCV potential threat to Solanaceae crops in China.The nearly full genomic sequence of LCV Shouguang city,Shandong province isolate(LCV-SD)was sequenced by RT-PCR plus conventional Sanger sequencing.The LCV-SD genomic sequence was compared,aligned with others isolates,and the phylogenetic tree constructed using MEGA 5.0.The results showed the sequence of RNA1 fragment of LCV-SD possesses the highest nucleotide identity(98.39%)with Chongqing isolate of China,lower nucleotide identity(84.27%~85.27%)with foreign isolates.The characteristics of sequence of RNA2 fragment of LCV-SD was similar as RNA1 fragment with the highest nucleotide identity(99.05%)with Chongqing isolate of China,and lower nucleotide identity(87.68%-88.36%)with foreign isolates.Phylogenetic analysis showed both fragments of LCV-SD was located in same cluster with isolates of other regions including Nanjing,Chongqing of China.CPm protein encoded by LCV was located on the surface of virion,is a effective target as antigen for making polyantibody.The CPm gene was prokaryotic expressed,and CPm protein was purified by affinity chromatography.The purified CPm protein was as antigen to immune New Zealand rabbit.The purified polyantibody concentration was up to 1.44 mg/mL,and could recognize LCV at diluting up to 1:64 000,and specific to LCV.The optimal dilution ratio was 10 000 for western blotting and Dot-ELISA detecting LCV.This polyantibody was an alternative and effective tool for LCVELISA,Western blotting and Dot-blotting detection.