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Titolo:
Expression of intron-containing GUS constructs is reduced due to activation of a cryptic 5 ' splice site
Autore:
Ibrahim, AFM; Watters, JA; Clark, GP; Thomas, CJR; Brown, JWS; Simpson, CG;
Indirizzi:
Scottish Crop Res Inst, Unit Gene Express, Dundee DD2 5DA, Scotland Scottish Crop Res Inst Dundee Scotland DD2 5DA Dundee DD2 5DA, Scotland Adv Technol Cambridge Ltd, Cambridge, England Adv Technol Cambridge Ltd Cambridge England dge Ltd, Cambridge, England
Titolo Testata:
MOLECULAR GENETICS AND GENOMICS
fascicolo: 3, volume: 265, anno: 2001,
pagine: 455 - 460
SICI:
1617-4615(200105)265:3<455:EOIGCI>2.0.ZU;2-O
Fonte:
ISI
Lingua:
ENG
Soggetto:
MESSENGER-RNA; HIGHER-PLANTS; 5'-SPLICE-SITE SELECTION; U1 SNRNA; SEQUENCES; MUTATION; PROTEIN; MARKER; NUCLEI; GENES;
Keywords:
GUS; intron; splicing; cryptic splicing; expression levels;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
23
Recensione:
Indirizzi per estratti:
Indirizzo: Simpson, CG Scottish Crop Res Inst, Unit Gene Express, Dundee DD2 5DA, Scotland Scottish Crop Res Inst Dundee Scotland DD2 5DA 5DA, Scotland
Citazione:
A.F.M. Ibrahim et al., "Expression of intron-containing GUS constructs is reduced due to activation of a cryptic 5 ' splice site", MOL GENET G, 265(3), 2001, pp. 455-460

Abstract

An intron-containing beta -glucuronidase (GUS) gene has been used widely in promoter analyses and as a plant transformation marker. Maximal plant gene expression requires accurate and efficient removal of the intron from theexpressed pre-mRNA transcripts by splicing. Detailed analysis of splicing of potato ST-LS1 and pea legumin introns from GUS constructs revealed the activation of a cryptic 5 ' splice site in the GUS coding sequence 4 nt upstream from the authentic intron 5 ' splice site. About 40% of transcripts utilised the cryptic 5 ' splice site in tobacco protoplasts, reducing the translational potential of expressed pre-mRNA. The same cryptic splicing eventwas evident in transgenic tobacco leaves but at reduced levels. Mutations that removed the cryptic 5 ' splice site are associated with a two-fold enhancement in GUS activity in tobacco protoplasts, highlighting the need for careful examination of introns and their sites of insertion into gene constructs to minimise variability in gene activity and maximise gene expression.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 25/09/20 alle ore 06:20:20