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针对烟草类囊体膜中与放氧复合体及 Rieske 铁硫蛋白相关组分的反义 RNA 可抑制 mRNA 的积累,但不抑制蛋白质的积累。

Antisense RNA for components associated with the oxygen-evolving complex and the Rieske iron/sulfur protein of the tobacco thylakoid membrane suppresses accumulation of mRNA, but not of protein.

作者信息

Palomares R, Herrmann R G, Oelmuller R

机构信息

Botanisches Institut der Ludwig-Maximilians-Universität, München, FRG.

出版信息

Planta. 1993;190(3):305-12. doi: 10.1007/BF00196958.

Abstract

Homologous cDNA clones of the nuclear-codes 23- and 33-kDa polypeptides of the oxygen-evolving system, as well as the Rieske iron/sulfur protein, were expressed in antisense or sense orientation under the control of the 35S RNA cauliflower mosaic virus promoter in transgenic tobacco (Nicotiana tabacum, L.) in order to modulate stationary concentrations of transcripts for studying relationships between transcript and protein levels. In all instances, in individuals that contained as little as 10% or less of the transcripts of untransformed plants, the corresponding protein levels were not notably altered. This indicated that the mRNA levels for components under study are not rate-limiting for protein accumulation and that severely reduced amounts of these transcripts still allow normal plant development. Overexpression of the 23-kDa polypeptide by the corresponding cDNA integrated in sense orientation resulted in both higher mRNA and protein levels without detectable enhancement of oxygen evolution. At least some of the excess protein was found in the soluble fraction and not associated with the photosystem-II reaction center. An attempt has been made to reconcile the varied responses obtained using these approaches with different expression patterns.

摘要

为了调节转录本的稳定浓度以研究转录本水平与蛋白质水平之间的关系,在转基因烟草(烟草,L.)中,在花椰菜花叶病毒35S RNA启动子的控制下,以反义或正义方向表达了放氧系统的23 kDa和33 kDa核编码多肽以及 Rieske 铁硫蛋白的同源cDNA克隆。在所有情况下,在含有低至未转化植物转录本10%或更少的个体中,相应的蛋白质水平没有明显改变。这表明所研究成分的mRNA水平不是蛋白质积累的限速因素,并且这些转录本的大量减少仍然允许植物正常发育。通过以正义方向整合的相应cDNA对23 kDa多肽进行过表达,导致mRNA和蛋白质水平均升高,但未检测到放氧增强。至少一些过量的蛋白质存在于可溶部分中,并且与光系统II反应中心无关。人们试图将使用这些方法获得的不同反应与不同的表达模式协调起来。

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