鸡蛋花花叶病毒3'端序列的克隆与分析

CLONING AND SEQUENCING OF THE 3'-END OF FRANGIPANI MOSAIC VIRUS (FMV)

摘要:

根据烟草花叶病毒组(Tobamoviruses)3'端非编码区保守区域,人工合成下游引物P1,P2,P3,分别以这些引物引导在反转录酶作用下合成双链cDNA,并克隆到pBS载体上。HindⅢ+PstI双酶切分析重组子后,得到15个阳性重组子,其中重组子pBF3含有2432 bp外源DNA。序列分析结果表明:该2432 bp的序列含2个开放读框,即鸡蛋花花叶病毒移动蛋白基因序列和外壳蛋白基因序列。移动蛋白基因序列起始于806位ATG,中止于l 576位TAG。推导的氨基酸序列共256个氨基酸,分子量为28.5 kDa。外壳蛋白基因序列起始于l 635位ATG,中止于2 158位TAA。推导的氨基酸序列共l74个氨基酸,分子量为19.4 kDa.此外该片段还包含部分180 kDa蛋白和全部3'端非编码区核苷酸序列。

Abstract:

Based on the 3'-end conserved regions among genomic RNA of tobamovirus, three primers P1, P2, P3 matching them were synthesized. ds-cDNA were synthesized to leading by these primers and cloned into the vector pBS. Fifteen positive recombinants were obtained after Hind Ⅲ + PstⅠ were digested. Among them, pBF3 was sequenced. It showed that the fragment of 2432 bp had two open reading frames (ORF). The first ORF encoded the frangipani mosaic virus movement protein. It ranged from nucleotide 806 to 1576. The deducing protein co nsisted of 256 amino acids．with a calculated MW of 28。5 kilo dalton．The second ORF encoded the coat protein which ranged from nuclco tide 1 635 to 2 158，and the deducing protein co nsisted of 1 74 amino acids，with a calculated MW of l9.4 kilo dalton． This frgment also included partial regions of 180 kDa protein and 3'-end non-coding regions．