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1. US20060286607 - Isolation of Proteins Involved in Posttranscriptional Gene Silencing and Methods of Use

Office
États-Unis d'Amérique
Numéro de la demande 11458933
Date de la demande 20.07.2006
Numéro de publication 20060286607
Date de publication 21.12.2006
Type de publication A1
CIB
C40B 30/06
CCHIMIE; MÉTALLURGIE
40TECHNOLOGIE COMBINATOIRE
BCHIMIE COMBINATOIRE; BIBLIOTHÈQUES, p.ex. CHIMIOTHÈQUES, BIBLIOTHÈQUES VIRTUELLES (IN SILICO)
30Procédés de criblage des bibliothèques
06en mesurant les effets sur des cellules, des tissus ou des organismes vivants
C07K 14/415
CCHIMIE; MÉTALLURGIE
07CHIMIE ORGANIQUE
KPEPTIDES
14Peptides ayant plus de 20 amino-acides; Gastrines; Somatostatines; Mélanotropines; Leurs dérivés
415provenant de végétaux
Déposants MIRKOV T E
INGELBRECHT IVAN L
Inventeurs Mirkov T. Erik
Ingelbrecht Ivan L.
Mandataires BAKER BOTTS L.L.P.
Données relatives à la priorité 10226715 23.08.2002 US
11252080 17.10.2005 US
60314863 24.08.2001 US
Titre
(EN) Isolation of Proteins Involved in Posttranscriptional Gene Silencing and Methods of Use
Abrégé
(EN)

The present invention includes a method for detecting and isolating sugarcane proteins that interact with the HC-Pro and P1 proteins of SrMV and other proteins involved in gene silencing, particularly in sugarcane. The method uses a two hybrid assay with an HC-Pro, P1, or other silencing-related protein-containing bait protein and a prey protein containing a polypeptide encoded by a DNA molecule in a cDNA library. The method also includes identification of false positives through reverse two-hybrid assays and using in vitro techniques such as farwestern blots or pull down assays where plant physiological conditions may be replicated. Finally, interactions may be confirmed in planta. Some novel proteins used in and discovered using the these methods are also identified. Methods of using viral and plant proteins to regulate silencing in plants such as sugarcane are also discussed.