[通过转录后基因沉默分析烟曲霉的 rbcS 基因功能]。

Xiao-Fu Zhou, Peng-Da Ma, Ren-Hou Wang, Xiao-Juan Zhu, Bao Liu, Xing-Zhi Wang
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引用次数: 0

摘要

以烟草鼠疫病毒载体和烟草烟碱为实验材料,建立并优化了病毒诱导转录后基因沉默系统,用于研究rbcS基因的功能。对沉默rbcS基因植株的表型特征、RT-PCR法检测rbcS基因转录水平、rbcS和rbcL抗体检测rbcS蛋白水平、HPLC法检测rbcS沉默植株光合色素含量进行了分析。结果表明,21-24 天的幼苗和农杆菌浓度为 OD600 = 1-1.5 的基因沉默效果最好。rbcL 的表达水平很可能受 rbcS 的调控,而 rbcS 基因与光合作用能量的收集无关。概率分析表明,烟草鼠疫病毒载体系统是通过转录后基因沉默研究 rbcS 基因功能的有效技术。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[The analysis of rbcS gene function by post-transcription gene silencing in Nicotiana benthamiana].

A system of virus-induced post-transcriptional gene silencing for studying rbcS gene function was established and optimized using tobacco rattle virus vector and Nicotiana benthamiana as experimental materiaes. The following analyses were conducted: phenotypic characterization of rbcS gene silenced plants, transcription levels of rbcS gene by RT-PCR; protein levels of rbcS by the antibodies of rbcS and rbcL and photosynthetic pigments wntents in rbcS silenced plants by HPLC method. The results showed that the seedlings at 21-24-day-old and Agrobacterium concentration at OD600 = 1-1.5 gave the best results for gene silencing. The expression level of rbcL was very likely regulated by rbcS, and rbcS gene did not relate to the collection of photosynthetic energy. Probability analysis showed that the tobacco rattle virus vector system is a useful and effective technique to study rbcS gene function via post-transcriptional gene silencing.

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