title: Development of new potato virus X-based vectors for gene over-expression and gene silencing assay.
abstract: Multiple plant viruses, including potato virus X (PVX), have been modified as vectors for expressing heterologous genes or silencing endogenous genes in plants. PVX-based vectors facilitate the functional analysis of genes in plant. However, they can only express one protein in a time. In this paper we report the construction of new vectors based on a 35S promoter-driven PVX infectious clone, pCaPVX100. Vector pCaPVX440 contains two additional subgenomic promoters and can be utilized to express two foreign genes at the same time. Plasmid pCaPVX760 is a CP minus vector and can be used to express foreign proteins through the gene substitution strategy. In addition, plasmid pCaPVX100 was engineered into a gene silencing vector (pCaPVX440-LIC) by introducing a ligation independent cloning (LIC) site into the vector. These results indicate that the newly developed PVX vectors are competent for multiple research purposes.
authors: Department of Plant Pathology, College of Plant Protection, Shandong Agricultural University, Shandong 271018, China; Department of Microbiology, College of Life Sciences, Shandong Agricultural University, Shandong 271018, China.
authors: Ying
authors: Y
authors: Wang
authors: Department of Plant Pathology, College of Plant Protection, Shandong Agricultural University, Shandong 271018, China.
authors: Qian-Qian
authors: QQ
authors: Cong
authors: Tai'an Academy of Agricultural Sciences, Shandong 271000, China.
authors: Yu-Fei
authors: YF
authors: Lan
authors: Department of Plant Pathology, College of Plant Protection, Shandong Agricultural University, Shandong 271018, China.
authors: Chao
authors: C
authors: Geng
authors: Department of Plant Pathology, College of Plant Protection, Shandong Agricultural University, Shandong 271018, China.
authors: Xian-Dao
authors: XD
authors: Li
authors: Department of Plant Pathology, College of Plant Protection, Shandong Agricultural University, Shandong 271018, China.
authors: Yuan-Cun
authors: YC
authors: Liang
authors: Department of Microbiology, College of Life Sciences, Shandong Agricultural University, Shandong 271018, China. Electronic address: zhyou21cn@sdau.edu.cn.
authors: Zheng-You
authors: ZY
authors: Yang
authors: Department of Plant Pathology, College of Plant Protection, Shandong Agricultural University, Shandong 271018, China.
authors: Xiao-Ping
authors: XP
authors: Zhu
authors: Department of Plant Pathology, College of Plant Protection, Shandong Agricultural University, Shandong 271018, China. Electronic address: xdongli@sdau.edu.cn.
authors: Xiang-Dong
authors: XD
authors: Li
mesh_terms: D003001:Cloning, Molecular; D015870:Gene Expression; D020868:Gene Silencing; D018390:Gene Targeting / Q000295:instrumentation*; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D010944:Plants / Q000235:genetics*; D017863:Potexvirus / Q000235:genetics* / Q000378:metabolism
keywords: Gene silencing; Gene vector; Ligation independent cloning; Over-expression; Potato virus X (PVX)