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内含子在转录后发挥作用,以增加拟南芥色氨酸途径基因PAT1的表达。

Introns act post-transcriptionally to increase expression of the Arabidopsis thaliana tryptophan pathway gene PAT1.

作者信息

Rose A B, Last R L

机构信息

Boyce Thompson Institute for Plant Research, Cornell University, Ithaca, NY 14853-1801, USA.

出版信息

Plant J. 1997 Mar;11(3):455-64. doi: 10.1046/j.1365-313x.1997.11030455.x.

Abstract

The expression of the Arabidopsis thaliana PAT1 gene, which encodes the tryptophan biosynthetic enzyme phosphoribosylanthranilate transferase, was investigated using translational fusions of the PAT1 promoter to the GUS reporter gene. Independent stably transformed A. thaliana lines containing a single copy of a fusion that includes the entire plastid transit peptide and the first two introns of PAT1 had on average 30 times more GUS enzyme activity than plants transformed with a construct in which GUS was fused a short distance downstream of the PAT1 start codon. Plants containing the construct without introns or leader peptide accumulated undetectable amounts of PAT1-GUS fusion protein and mRNA, even though the transcriptional rate of both fusion constructs was comparable. Fusions containing the entire transit peptide and either of the first two introns yield as much GUS activity as constructs containing both introns, but constructs containing the transit peptide but no introns give rise to much lower levels. Therefore, introns greatly enhance the expression of PAT1-GUS fusions, and they act post-transcriptionally to increase the steady-state level of mRNA.

摘要

利用拟南芥PAT1启动子与GUS报告基因的翻译融合,研究了编码色氨酸生物合成酶磷酸核糖基邻氨基苯甲酸转移酶的拟南芥PAT1基因的表达情况。独立稳定转化的拟南芥株系含有一个融合基因的单拷贝,该融合基因包括整个质体转运肽和PAT1的前两个内含子,其GUS酶活性平均比用GUS融合在PAT1起始密码子下游短距离处的构建体转化的植物高30倍。不含内含子或前导肽的构建体的植物,即使两种融合构建体的转录速率相当,也积累了无法检测到的PAT1-GUS融合蛋白和mRNA。含有整个转运肽和前两个内含子之一的融合体产生的GUS活性与含有两个内含子的构建体一样多,但含有转运肽但不含内含子的构建体产生的水平要低得多。因此,内含子极大地增强了PAT1-GUS融合体的表达,并且它们在转录后起作用以增加mRNA的稳态水平。

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