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Silencing of potato virus X coat protein gene in transgenic tobaccos by codon replacement that confers resistance to PVX infection
Authors:Feng  Dejiang  Liu  Xiang  Meng  Kun  Liao  Lili  Wei  Xiaoli  Xu  Honglin  Zhu  Zhen
Affiliation:e-mail: zzhu@ genetics.ac.cn
Abstract:To understand the effect of rare codon on the silencing ratio of foreign gene, some preferred codon in potato virus X (PVX) coat protein gene (cp) were substituted with synonymous rare codons. The modified PVX coat protein gene (cpm) and wild-type cp gene (cpw) were inserted into binary vector under the control of CaMV35S promoter, and these two plant expression constructs were transferred into tobacco (Nicotiana tabacum cv. Xanthi) genomes via Agrobacterium mediated method and transgenic plants were generated. Northern blot analysis of RNA isolated from these plants showed that the silencing ratio of cpm gene in transgenic tobaccos was higher than that of cpw (35% and 6.25% respectively). Run on results indicate that the silencing of cp gene happened at post-transcriptional level. The resistance of transgenic tobaccos carrying cpm genes to PVX is increased compared with that of transformants carrying cpw genes. These results suggest that the resistance of transgenic tobacco to PVX can be enhanced by codon replacement.
Keywords:potato virus X   coat protein   codon modification   gene silencing   virus resistance.
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