Sequencing and construction of plant expression vector of coat protein
gene from Ys strain of pa- paya ringspot virus

YE Chang-Meng; FAN Huai-Zhong; YE Yin; TIAN Bo

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February 1997

YE Chang-Meng, FAN Huai-Zhong, YE Yin and TIAN Bo. Sequencing and construction of plant expression vector of coat proteingene from Ys strain of pa- paya ringspot virus[J]. Virologica Sinica, 1997, 12(1).

Citation: YE Chang-Meng, FAN Huai-Zhong, YE Yin, TIAN Bo. Sequencing and construction of plant expression vector of coat protein gene from Ys strain of pa- paya ringspot virus .VIROLOGICA SINICA, 1997, 12(1) : 54.

番木瓜环斑病毒Ys CP基因的序列分析和植物表达载体的构建

1.
中国科学院微生物研究所华南农业大学植保系中山大学生物工程研究中心工作

摘要

用双脱氧链终止法分析了克隆的番木瓜环斑病毒（PRV）Ys株系外壳蛋白（CP）基因的序列，结果表明YsCP基因全长858nt。对PRV国内外16个株系或分离物CP基因的比较发现，YsCP基因与国内株系或分离物CP基因的同源性较高（94．44％～97．68％），而与国外株系或分离物CP基因的同源性较低（88．88％～92．70％）。CP基因之间的差异主要靠近基因5’端，特别是在YSCP基因第63nt后连续缺失6nt，SmGTHAI和SRI的CP基因也在此处缺失3nt。将YsCP基因插入中间质粒pRokⅡ的CaMV35S启动于和nos终止序列之间形成CP基因的植物表达载体pRPCY，通过三亲交配使pRPCY进入农杆菌LBA4404，与其中的pAL4404构成双元载体系统。
- 番木瓜环斑病毒外壳蛋白基因植物表达载体

Sequencing and construction of plant expression vector of coat protein gene from Ys strain of pa- paya ringspot virus

Abstract

The cloned coat protein (CP) gene from Ys strain of papaya rinspot virus (PRV) was sequenced by dideoxy-chain termination method. Results show that the full length of the CP genewas 858 nucleotides (nt). Comparison among CP genes of 16 PRV strains or isolates from Chinaand abroad indicated that Ys shared high CP gene homologous sequence (94.44%~97.68%)with the other strains or isolates from China, while it shared lower CP gene homologous sequence(88.88%-92.70% )with the strains or isolates abroad. It was noted that there were 6 nt missing after the 63rd nt in Ys CP gene, and there were 3 nt missing in CP genes of Sin. G. THAI.SRI. Ys CP cDNA was inserted into the cloning sites between CaMV 35S promoter and nos terminator of the middle plasmid pRok Ⅱ to produce the plant expression vector pRPCY. LBA44004of Agrtobacterium tumefasciens containing the binary vector system pRPCY/pAL4404 was obtained through triparental mating, and will be used to introduce CP gene into papaya genome.
- Papaya ringspot virus Coat protein gene Plant expression vector

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Sequencing and construction of plant expression vector of coat protein gene from Ys strain of pa- paya ringspot virus

Abstract: The cloned coat protein (CP) gene from Ys strain of papaya rinspot virus (PRV) was sequenced by dideoxy-chain termination method. Results show that the full length of the CP genewas 858 nucleotides (nt). Comparison among CP genes of 16 PRV strains or isolates from Chinaand abroad indicated that Ys shared high CP gene homologous sequence (94.44%~97.68%)with the other strains or isolates from China, while it shared lower CP gene homologous sequence(88.88%-92.70% )with the strains or isolates abroad. It was noted that there were 6 nt missing after the 63rd nt in Ys CP gene, and there were 3 nt missing in CP genes of Sin. G. THAI.SRI. Ys CP cDNA was inserted into the cloning sites between CaMV 35S promoter and nos terminator of the middle plasmid pRok Ⅱ to produce the plant expression vector pRPCY. LBA44004of Agrtobacterium tumefasciens containing the binary vector system pRPCY/pAL4404 was obtained through triparental mating, and will be used to introduce CP gene into papaya genome.