关键词: 柑橘黄化脉明病毒, RACE, 全长cDNA, 长链RT-PCR

Abstract:

In order to clone full-length cDNA of Citrus yellow vein clearing virus（CYVCV），further understand the molecular characteristics of CYVCV and its pathogenic mechanism, a long RT-PCR system was established with optimizing conditions and specific primer designed according to the sequences of CYVCV in GenBank and the results of 5′RACE（rapidamplification of cDNA ends）. After extraction of total RNA from the Citrus aurantium‘Daidai’of the infected CYVCV and amplification the full-length genome of CYVCV by the long RT-PCR，the full-length of CYVCV cDNA was obtained and cloned into pGEM-T easy vector and sequenced. The results showed that the 7.5 kb full-length genome of Citrus yellow vein clearing virus was amplified successfully by the long RT-PCR. The homology of nucleotide sequences of the four amplified full-length cDNA were 93%–99% to the strains in GenBank.

Key words: Citrus yellow vein clearing virus（CYVCV）, RACE, full-length cDNA, long RT-PCR