CAJ | 학술논문

XJ-160 virus was isolated in 1990 from Xinjiang, China. Thebiological characteristics of XJ-160 virus suggested that it might be a Togavirus. The serological tests showed that XJ-160 virus is a Sindbis-like virus. The complete 11626-base nucleotide sequence of XJ-160 has been determined recently.It is necessary to establish a reverse genetic system for rescue of XJ-160 virus from its cDNA clone. We describe here the constructon of full-length cDNA clone of XJ-160 virus.The total RNA were extracted from BHK cells infected with XJ-160 virus. Specific primers were used to amplify five fragments covering the whole genome of XJ-160 virus. All of these five fragments have the restriction enzyme sites at both termini to be assembled into the full-length cDNA clone. The five PCR fragments were cloned into pGEM-T vector. The clones with SP6-inserted direction were selected for subsequent manipulation. After a series of plasmid manipulation, the full-length cDNA clone of XJ-160 virus was assembled into pBluescript vector. The restriction enzyme assay and sequencing analysis demonstrated that the whole genome of XJ-160 virus was correctly assembled into pBluescript vector, with added SP6 promoter in 5′ terminus and poly A in its 3′ terminus.