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[马铃薯X病毒5'前导序列对转基因马铃薯中马铃薯Y病毒膜蛋白基因表达的影响]

[The effect of the 5'-leader of potato X-virus on expression of the gene for the potato Y virus membrane protein in transgenic Solanum tuberosum].

作者信息

Pugin M M, Sokolova M A, Shul'ga O A, Skriabin K G

出版信息

Mol Biol (Mosk). 1994 Jul-Aug;28(4):752-60.

PMID:7990802
Abstract

The 5'-leader of potato virus X (PVX) genomic RNA was successfully used to generate potato plants expressing the coat protein of the potato virus YN (Russian isolate) (PVY CP). Two expression cassettes were constructed that carried the PVY CP coding sequence along with the PVY 3'untranslated region under the control of the CaMV 35S promoter and poly(A) signal. The cassettes contained different sequences between the 35S promoter and the CP artificial AUG initiating codon of the coat protein: either a polylinker or a shortened variant of the PVX leader. Transgenic Belorusskii-3 potato plants generated by agrobacterial transformation were shown to express the PVY CP gene to different extents depending on the cassette integrated into the plant genome. The cassette containing the PVX leader provided detectable amounts of the coat protein in all tested plants, whereas plants carrying the cassette with the polylinker leader produced no coat protein above the Western detection threshold. Northern analysis revealed that the presence of the PVX leader in front of the PVY CP gene initiating codon resulted in a 10-fold increase in the PVY CP mRNA steady-state levels, thus suggesting an influence of the PVX leader on mRNA stability.

摘要

马铃薯X病毒(PVX)基因组RNA的5′前导序列成功用于培育表达马铃薯Y病毒N株系(俄罗斯分离株)(PVY CP)外壳蛋白的马铃薯植株。构建了两个表达盒,它们携带PVY CP编码序列以及CaMV 35S启动子和多聚腺苷酸信号控制下的PVY 3′非翻译区。这些表达盒在35S启动子和外壳蛋白的CP人工AUG起始密码子之间包含不同序列:一个多克隆位点或PVX前导序列的缩短变体。通过农杆菌转化产生的转基因别洛鲁斯基-3马铃薯植株根据整合到植物基因组中的表达盒不同程度地表达PVY CP基因。含有PVX前导序列的表达盒在所有测试植株中都能检测到外壳蛋白,而携带含有多克隆位点前导序列表达盒的植株在Western检测阈值以上未产生外壳蛋白。Northern分析表明,PVY CP基因起始密码子前存在PVX前导序列导致PVY CP mRNA稳态水平提高了10倍,因此表明PVX前导序列对mRNA稳定性有影响。

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