英语翻译RESULTSThe genomic structure of SPL3,SPL4 and SPL5cDNA sequences deposited in GenBank,as well as data from theTiling Array Transcriptome Express Tool (Yamada et al.,2003),indicate that SPL3 (At2g33810) has a complex transcription pattern.

英语翻译RESULTSThe genomic structure of SPL3,SPL4 and SPL5cDNA sequences deposited in GenBank,as well as data from theTiling Array Transcriptome Express Tool (Yamada et al.,2003),indicate that SPL3 (At2g33810) has a complex transcription pattern.
英语翻译
RESULTS
The genomic structure of SPL3,SPL4 and SPL5
cDNA sequences deposited in GenBank,as well as data from the
Tiling Array Transcriptome Express Tool (Yamada et al.,2003),
indicate that SPL3 (At2g33810) has a complex transcription pattern.
To determine the structure of this locus,we performed and
RACE in both sense and antisense orientations.These experiments
revealed that the SPL3 sense transcript (ST) has a single end and
multiple poly(A) sites in its UTR (Fig.1A).The miR156 target
site is located in the UTR,50 nucleotides from the stop codon.We
also identified several 950 nucleotide antisense transcripts (AST),
which completely encompass the ST and have an intron located in
nearly the same position as the intron in the ST (Fig.1A).Four
transcription start sites and 5 poly(A) sites were identified in these
AS transcripts.
SPL4 (At1g53160) and SPL5 (At3g15270) encode,respectively,
174 amino acid and 181 amino acid proteins that are 65% identical
(76% similar) to the 131 amino acid protein encoded by SPL3.The
structure of these genes is also very similar to SPL3 (Fig.1A).These
genes also possess a miR156 target site located within their UTR
and RLM-RACE demonstrated that SPL3/4/5 are all cleaved in the
middle of this target site (Fig.1B)
The regulation and function of SPL3
In order to study the role of miR156 in the regulation of SPL3,we
produced transgenic plants expressing miR156-sensitive and
miR156-resistant forms of this gene under the regulation of the
constitutive CaMV 35S promoter.We chose to use the 35S promoterrather than the endogenous SPL3 promoter in order to eliminate
effects on SPL3 expression due to transcriptional cross-regulation.
The constructs generated for this experiment express an SPL3
transcript with a normal 3\2 UTR (35S::SPL3),a transcript with seven
mutations in the miR156 target site (35S::SPL3m),and a transcript
lacking the 3\2 UTR (35S::SPL3\5) (Fig.2A).Constructs in which
the GUS Plus sequence was fused in frame to the 5\2 end of these
sequences were also generated in order to evaluate the expression of
the SPL3 protein.Plants expressing GUS-tagged versions of SPL3
resembled plants expressing untagged proteins,although their
phenotype was slightly weaker.Homozygous stocks containing a
single insert were established from transgenic lines,and these stocks
were subjected to Northern analysis to ensure that the transgene was
overexpressed.

英语翻译RESULTSThe genomic structure of SPL3,SPL4 and SPL5cDNA sequences deposited in GenBank,as well as data from theTiling Array Transcriptome Express Tool (Yamada et al.,2003),indicate that SPL3 (At2g33810) has a complex transcription pattern.
结果基因结构spl3,并spl5spl4存入同源基因序列, 以及数据块从阵列转录表达工具(山田等. ,2003), 显示spl3(at2g33810)具有复杂的转录模式. 确定这一轨迹结构,我们演出和3~5~两个种族意识和反方向. 这些实验显示spl3感全文(路段)已结束,多单5~6聚(一)第3点~翻译 (图一A) 该址位于mir156靶3~翻译,来自50个核苷酸终止密码. 我们还发现几个~950核苷酸反转录酶(AST) 其中,有一个完全包含圣内含位于差不多同一位置的内含在圣 (图一A) 4、5点开始转录聚(一)发现这些地点作为誊本. spl4(at1g53160)spl5(at3g15270)编码,分别 174、181个氨基酸的蛋白质胺基酸是65%相同(类似76%)的131个氨基酸蛋白质恩 由spl3编码. 这些基因的结构也很相似spl3(图1A)条. 这些基因还拥有自己mir156靶址位于3、5~翻译~RLM-RACE法都显示spl3/4/5 在这个目标中把整工地(图1B)款规和功能spl3为了研究 mir156作用的调节spl3, 我们生产的转基因植物表达mir156敏感和mir156西林形式下调控这个基因序列基本构 启动. 我们选择使用的35S启动promoterrather比源性spl3为了消除影响表达spl3 由于两岸转录调控. 这个实验所产生的建构,表达了与正常3spl3全文翻译(的35S::spl3) 誊7mir156突变的靶点(的35S::spl3m) 3、缺乏翻译全文(的35S::spl3)(图2A)条. 施工中的GUS+序列帧融合到5月底,在这些序列也 为了评价产生的spl3蛋白表达. 植物表达的GUS标记版本未贴spl3近似植物蛋白表达,其表型虽稍减弱. 合子插入建立单一股票含转基因线 这些股票受到北方分析,以确保被过度转.