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Titolo:
Identification of two short internal ribosome entry sites selected from libraries of random oligonucleotides
Autore:
Owens, GC; Chappell, SA; Mauro, VP; Edelman, GM;
Indirizzi:
Inst Neurosci, San Diego, CA 92121 USA Inst Neurosci San Diego CA USA 92121 st Neurosci, San Diego, CA 92121 USA Scripps Clin & Res Inst, Dept Neurobiol, La Jolla, CA 92037 USA Scripps Clin & Res Inst La Jolla CA USA 92037 iol, La Jolla, CA 92037 USA Skaggs Inst Chem Biol, La Jolla, CA 92037 USA Skaggs Inst Chem Biol La Jolla CA USA 92037 Biol, La Jolla, CA 92037 USA
Titolo Testata:
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
fascicolo: 4, volume: 98, anno: 2001,
pagine: 1471 - 1476
SICI:
0027-8424(20010213)98:4<1471:IOTSIR>2.0.ZU;2-W
Fonte:
ISI
Lingua:
ENG
Soggetto:
GREEN FLUORESCENT PROTEINS; FACTOR MESSENGER-RNA; 5'-UNTRANSLATED REGION; TRANSLATION INITIATION; BINDING; COMPLEMENTARITY; ELEMENTS; SEGMENT; CELLS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
24
Recensione:
Indirizzi per estratti:
Indirizzo: Owens, GC Inst Neurosci, 10640 John Jay Hopkins Dr, San Diego, CA 92121 USA Inst Neurosci 10640 John Jay Hopkins Dr San Diego CA USA 92121 A
Citazione:
G.C. Owens et al., "Identification of two short internal ribosome entry sites selected from libraries of random oligonucleotides", P NAS US, 98(4), 2001, pp. 1471-1476

Abstract

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 sequenceof the mRNA encoding Gtx homeodomain protein could function as an IRES, Toidentify 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, tells 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 I in ked 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 identifyIRES modules with specific expression properties may be useful in the design of vectors for biotechnology and gene therapy.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 01/12/20 alle ore 22:30:03