Identification of two short internal ribosome entry sites selected from libraries of random oligonucleotides.
Identification of two short internal ribosome entry sites selected from libraries of random oligonucleotides.
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鉴定从随机寡核苷酸文库中选择的两个短内部核糖体进入位点。
DOI：
10.1073/pnas.98.4.1471
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发表时间：
2001
影响因子：
11.1
通讯作者：
Edelman,GM
中科院分区：
文献类型：
--
作者：
Owens,GC;Chappell,SA;Mauro,VP;Edelman,GM
控制mRNA翻译的序列可能在调节蛋白质水平中起关键作用。一个这样的元件是内部核糖体进入位点（IRES）。我们以前的研究表明，Gtx同源结构域蛋白mRNA 5′前导序列中的一个9-nt片段可以作为IRES发挥功能。为了鉴定具有类似性质的其他短序列，我们设计了一种选择程序，该程序使用逆转录病毒载体来表达编码增强的绿色和青色荧光蛋白的双顺反子mRNA，分别作为第一顺反子和第二顺反子。 第二个顺反子的表达依赖于顺反子间序列，并指示IRES活性。B104细胞用两个逆转录病毒文库感染，所述逆转录病毒文库在顺反子间区域中含有9或18 nt的随机序列。对表达两种顺反子的细胞进行分选，并对从所选细胞回收的序列在双荧光素酶双顺反子mRNA中的IRES活性进行再分析。通过该程序鉴定了两种新型IRES，并且都含有与18 S rRNA互补的片段。当任一片段的多个拷贝连接在一起时，IRES活性显著增强。 此外，这些合成的IRES在各种细胞类型中具有不同的活性。这些特性与先前鉴定的来自Gtx mRNA的9-nt IRES模块的特性相似。这些结果提供了进一步的证据，短核苷酸序列可以作为IRES和支持的想法，一些细胞IRES可能是由较短的功能模块。鉴定具有特定表达特性的IRES模块的能力可用于设计用于生物技术和基因治疗的载体。
Sequences that control translation of mRNA may play critical roles in regulating protein levels. One such element is the internal ribosome entry site (IRES). We previously showed that a 9-nt segment in the 5′ leader sequence of the mRNA encoding Gtx homeodomain protein could function as an IRES. To identify other short sequences with similar properties, we designed a selection procedure that uses a retroviral vector to express dicistronic mRNAs encoding enhanced green and cyan fluorescent proteins as the first and second cistrons, respectively. Expression of the second cistron was dependent upon the intercistronic sequences and was indicative of IRES activity. B104 cells were infected with two retroviral libraries that contained random sequences of 9 or 18 nt in the intercistronic region. Cells expressing both cistrons were sorted, and sequences recovered from selected cells were reassayed for IRES activity in a dual luciferase dicistronic mRNA. Two novel IRESes were identified by this procedure, and both contained segments with complementarity to 18S rRNA. When multiple copies of either segment were linked together, IRES activities were dramatically enhanced. Moreover, these synthetic IRESes were differentially active in various cell types. These properties are similar to those of the previously identified 9-nt IRES module from Gtx mRNA. These results provide further evidence that short nucleotide sequences can function as IRESes and support the idea that some cellular IRESes may be composed of shorter functional modules. The ability to identify IRES modules with specific expression properties may be useful in the design of vectors for biotechnology and gene therapy.
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影响因子：
11.4
作者：
NIELSEN, PJ;TRACHSEL, H
通讯作者：
TRACHSEL, H
基因表达的翻译控制，由 Nahum Sonenberg、John W.B. 编辑
影响因子：
4.5
作者：
V. M. Pain;N. Standart
通讯作者：
N. Standart
DOI：
10.1083/jcb.150.1.275
发表时间：
2000-07-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Créancier L;Morello D;Mercier P;Prats AC
通讯作者：
Prats AC
通过核苷酸序列变异和活性选择获得的合成启动子元件。
DOI：
10.1073/pnas.97.7.3038
发表时间：
2000
影响因子：
11.1
作者：
G. Edelman;R. Meech;G. Owens;F. S. Jones
通讯作者：
F. S. Jones
用含有三顺反子 IRES 的逆转录病毒载体重建代谢途径以纠正四氢生物蝶呤缺陷
影响因子：
3.5
作者：
S. Laufs;Seon Hee Kim;Sunyoung Kim;N. Blau;B. Thöny
通讯作者：
B. Thöny