Citrus Phloem Transcriptional Profiling Using a Citrus Tristeza Virus Vectored Translating Ribosome Affinity Purification System

A phloem-limited citrus tristeza virus (CTV) vector was developed for the expression of an immune-tagged ribosomal protein, hfRPL18. CTV expressed hfRPL18 was used to pull down phloem derived polysomes from infected Citrus macrophylla leaf tissues. Associated hfRPL18 mRNAs were subsequently isolated and sequenced. To remove background mRNAs, isolated mRNAs from leaf tissues infected with the empty CTV vector were similarly sequenced. In summary, the CTV-hfRPL18 vector provides a transgene-free means to examine translating mRNAs within citrus phloem tissues.

一种用于表达免疫标记的核糖体蛋白hfRPL18的柑橘树斑萎病毒（CTV）传毒载体已被构建。通过CTV表达的hfRPL18，从感染了柑橘大叶的叶片组织中的韧皮部分离的多聚核糖体中被拉取。随后，相关的hfRPL18 mRNA被分离并测序。为了去除背景mRNA，同样对感染了空CTV传毒载体的叶片组织中的mRNA进行了测序。总之，CTV-hfRPL18传毒载体提供了一种无需转基因的方法，用于研究柑橘韧皮部组织中的翻译mRNA。