University of Illinois at Urbana-Champaign
Structure of the amplified 5-enolpyruvylshikimate-3-phosphate synthase gene in glyphosate-resistant carrot cells and attempts to isolate the anthranilate synthase gene from plants
Abstract
dc:descriptionAnalysis of the structure of amplified 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) DNA of carrot suspension cultured cell lines selected for glyphosate resistance was carried out to determine the mechanism of gene amplification in this plant system. Cloning of the EPSPS gene and 5$\sp\prime$ flanking sequences was carried out and two different DNA structures were revealed. The 13 kbp clone contained only one copy of the EPSPS gene while the 16 kbp clone contained an inverted duplication of the gene. Southern blot analysis with cloned carrot DNA as a probe showed that only the non-inverted repeated DNA structure was present in all of the cell lines during the selection process and the inverted repeat (IR) was present only in highly amplified DNA. The two structures were present in about equal numbers in the highly amplified line, TC 35G. The presence of the IR was further verified by resistance to Sl nuclease hydrolysis after denaturation and rapid renaturation, showing foldback DNA. This is the first report of an IR in amplified DNA of a target enzyme gene in selected plant cells.
Degree
thesis:*- Name thesis:degree_name
- Ph.D.
- Level thesis:degree_level
- Dissertation
- Discipline thesis:degree_discipline
- Biology
- Grantor
- University of Illinois at Urbana-Champaign
- Year dc:date
- 2011
Author and committee
dc:creator, dc:contributor.*- Author dc:creator
-
- Suh, Hyang
- Contributors dc:contributor
-
- Widholm, Jack M.
Subjects
dc:subject × 3Rights
dc:rights- Statement dc:rights
-
- Copyright 1992 Suh, Hyang
- Language dc:language
- eng
Identifiers
dc:identifier.*- Identifier
-
AAI9215891
(UMI)AAI9215891 - OAI identifier oai:identifier
- oai:www.ideals.illinois.edu:2142/22417