英语翻译The GFP-RTE1 Fusion Protein Is Localized to the Golgiand

英语翻译
The GFP-RTE1 Fusion Protein Is Localized to the Golgi
and Causes Ethylene Insensitivity
In an attempt to identify the subcellular localization
of the RTE1 protein,each of the RTE1 N and C termini
was fused with the GFP and ectopically expressed
under the control of the native RTE1 or 35S promoter.
The RTE1-GFP fusion,of which the RTE1 C terminus
was fused with GFP,failed to cause ethylene
insensitivity in wild type and to rescue the rte1-2
mutation in rte1-2 etr1-2 (data not shown).The GFPRTE1
fusion,of which GFP was fused to the N terminus
of RTE1,was able to cause some degrees of
ethylene insensitivity in wild type and partially rescue
the rte1-2 mutation in rte1-2 etr1-2 (Fig.6,A and B).The
expression of 35STGFP-rte1-2 failed to,respectively,
restore and confer ethylene insensitivity in etr1-2 rte1-2
and wild type (data not shown).
Fluorescence patterns of 35STGFP-RTE1- and
RTE1pTGFP-RTE1-transformed wild type were similar
to that of the 35STGFP-rte1-2 transformed,except
that RTE1pTGFP-RTE1 gave a weaker fluorescence
signal (see Supplemental Fig.S1).These results indicate
that the GFP-RTE1 fusion protein is functional
and dominant,and that its subcellular localization
could be sites where RTE1 exerts its function.Besides,
the rte1-2 mutation may cause dysfunction of RTE1,
rather than mislocalization as previously proposed
(Resnick et al.,2006).
Laser scanning confocal microscopy (LSCM) shows
that the green fluorescence signal of GFP-RTE1 was
fast moving,suggesting that its movement is associated
with cellular structures in the cytoplasm (Fig.6C).
Because RTE1 encodes a membrane protein,we next
examined if RTE1 may associate with any endomembranes,
including the ER and Golgi.GFP-ER,an
ER-localized marker,displayed a unique reticular fluorescence
pattern that is distinct to GFP-RTE1 and EYFPNAG
(see Supplemental Fig.S2),of which EYFP-NAG
was a marker for the Golgi localization (Xu and Scheres,
2005).When GFP-RTE1 and the Golgi-localized EYFPNAG
were coexpressed in onion epidermal cells,both
GFP-RTE1 and EYFP-NAG displayed fast movement
and the fluorescence visually overlapped (Fig.6D).
内容来自RTE1 Is a Golgi-Associated and ETR1-Dependent Negative Regulator of Ethylene Responses
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绿色荧光蛋白RTE1融合蛋白是本地化的高尔基体和乙烯不敏感的原因.在设法确定亚细胞定位的RTE1蛋白质,每个RTE1 N和C端融合的绿色荧光蛋白和ectopically表示控制下的本土RTE1或35S启动.该RTE1 - GFP融合,其中RTE1 ç总站是融合蛋白,但并未造成乙烯敏感的野生型和抢救rte1 - 2基因突变的rte1 - 2 etr1 - 2 （数据未显示） .该GFPRTE1融合,其中绿色荧光蛋白融合到N端RTE1 ,能够造成一定程度的乙烯敏感的野生型和部分抢救rte1 - 2基因突变的rte1 - 2 etr1 - 2 （图6 ,A和B ） .表达35STGFP - rte1 - 2没有分别,恢复和授予乙烯敏感的etr1 - 2 rte1 - 2和野生型（数据未显示） .
荧光模式35STGFP - RTE1和RTE1pTGFP - RTE1转化的野生型相似,在35STGFP - rte1 - 2转变,但RTE1pTGFP - RTE1了较弱的荧光信号（见补充图.一） .这些结果表明,绿色荧光蛋白RTE1融合蛋白的功能和优势,其亚细胞定位可以地点RTE1发挥其功能.此外,rte1 - 2突变可能会导致功能障碍的RTE1 ,不是mislocalization先前提议（ Resnick先生等人.,2006年） .
激光扫描共聚焦显微镜（激光共聚焦显微镜）表明,绿色荧光信号的绿色荧光蛋白RTE1是快速移动,这意味着其运动与细胞结构中的细胞质（图6C条） .
由于RTE1编码一个膜蛋白,我们下次审查,如果准RTE1可能与任何endomembranes ,包括ER和高尔基.急诊本地化标记,显示了独特的网状荧光模式是独特的,以绿色荧光蛋白RTE1和EYFPNAG （见补充图.S2 ） ,其中EYFP葡萄糖苷是一种标记的高尔基本地化（徐和Scheres ,2005年） .当绿色荧光蛋白RTE1和高尔基本地化EYFPNAG被coexpressed在洋葱表皮细胞,两个绿色荧光蛋白RTE1和EYFP葡萄糖苷显示快速运动和荧光视觉重叠（图六自由度） .
内容来自RTE1是一个高尔基相关和ETR1变负调控乙烯反应