RUcore: Rutgers University Community Repository
Site-specific recombinases to manipulate the plastid genome
Descriptive
TypeOfResource
Text
TitleInfo
(ID = T-1);
(type = uniform)
Title
Site-specific recombinases to manipulate the plastid genome
Identifier
(type = hdl)
http://hdl.rutgers.edu/1782.2/rucore10001600001.ETD.16733
Identifier
ETD_429
Language
LanguageTerm
(authority = ISO639-2)
eng
Genre
(authority = marcgt)
theses
Subject
(ID = SBJ-1);
(authority = RUETD)
Topic
Plant Biology
Subject
(ID = SBJ-1);
(authority = ETD-LCSH)
Topic
Plastids
Subject
(ID = SBJ-1);
(authority = ETD-LCSH)
Topic
Plant genetics
Subject
(ID = SBJ-1);
(authority = ETD-LCSH)
Topic
Genetic recombination
Subject
(ID = SBJ-1);
(authority = ETD-LCSH)
Topic
Recombinant DNA
Abstract
The plastid genome (ptDNA) of higher plants is highly polyploid and the 1,000 to 10,000 copies are compartmentalized in up to ~100 plastids per cell. I report here two different site-specific recombinases to manipulate the tobacco plastid genome, the P1 phage Cre recombinase and the phiC31 phage Integrase (Int). Thus far plastid transformation in higher plants was based on incorporation of foreign DNA in the plastid genome by the plastid's homologous recombination machinery. I describe here an alternative approach that relies on integration of foreign DNA by Int mediating recombination between bacterial attB and phage attP sequences. During the plastid transformation protocol marker genes are essential for selective amplification of rare transformed plastid genome copies to obtain genetically stable transplastomic plants. However, the marker gene becomes dispensable once homoplastomic plants are obtained. I describe plastid marker gene excision with both the Cre-loxP and the Int-att site-specific recombinases. The Cre-loxP approach uses a transiently expressed Cre, a novel protocol that enables rapid removal of marker genes from the ~10,000 plastid genome copies without transformation of the plant nucleus. The Int-att system uses a nuclear-encoded plastid-targeted Int to efficiently excise plastid marker genes flanked by directly oriented attB and attP sites. Lastly, I describe use of the Int-att system to study plastid sorting with the conclusion that a newly arising minor plastid DNA type can get established in a developing tobacco shoot and transmitted to the seed progeny.
PhysicalDescription
Extent
xii, 211 pages
InternetMediaType
application/pdf
InternetMediaType
text/xml
Note
(type = degree)
Ph.D.
Note
(type = bibliography)
Includes bibliographical references.
Name
(ID = NAME-1);
(type = personal)
NamePart
(type = family)
Lutz
NamePart
(type = given)
Kerry A.
Role
RoleTerm
(authority = RULIB);
(type = )
author
DisplayForm
Kerry A. Lutz
Name
(ID = NAME-2);
(type = personal)
NamePart
(type = family)
Maliga
NamePart
(type = given)
Pal
Role
RoleTerm
(authority = RULIB);
(type = )
chair
Affiliation
Advisory Committee
DisplayForm
Pal Maliga
Name
(ID = NAME-3);
(type = personal)
NamePart
(type = family)
Dooner
NamePart
(type = given)
Hugo
Role
RoleTerm
(authority = RULIB);
(type = )
internal member
Affiliation
Advisory Committee
DisplayForm
Hugo Dooner
Name
(ID = NAME-4);
(type = personal)
NamePart
(type = family)
Kerstetter
NamePart
(type = given)
Randall
Role
RoleTerm
(authority = RULIB);
(type = )
internal member
Affiliation
Advisory Committee
DisplayForm
Randall Kerstetter
Name
(ID = NAME-5);
(type = personal)
NamePart
(type = family)
Carrer
NamePart
(type = given)
Helaine
Role
RoleTerm
(authority = RULIB);
(type = )
outside member
Affiliation
Advisory Committee
DisplayForm
Helaine Carrer
Name
(ID = NAME-6);
(type = personal)
NamePart
(type = family)
Rongo
NamePart
(type = given)
Christopher
Role
RoleTerm
(authority = RULIB);
(type = )
outside member
Affiliation
Advisory Committee
DisplayForm
Christopher Rongo
Name
(ID = NAME-1);
(type = corporate)
NamePart
Rutgers University
Role
RoleTerm
(authority = RULIB);
(type = )
degree grantor
Name
(ID = NAME-2);
(type = corporate)
NamePart
Graduate School - New Brunswick
Role
RoleTerm
(authority = RULIB);
(type = )
school
OriginInfo
DateCreated
(point = );
(qualifier = exact)
2007
DateOther
(qualifier = exact);
(type = degree)
2007
Location
PhysicalLocation
(authority = marcorg)
NjNbRU
RelatedItem
(type = host)
TitleInfo
Title
Graduate School - New Brunswick Electronic Theses and Dissertations
Identifier
(type = local)
rucore19991600001
Identifier
(type = doi)
doi:10.7282/T3Q81DHF
Genre
(authority = ExL-Esploro)
ETD doctoral
Back to the top
Rights
RightsDeclaration
(AUTHORITY = GS);
(ID = rulibRdec0006)
The author owns the copyright to this work.
Copyright
Status
Copyright protected
Availability
Status
Open
AssociatedEntity
(AUTHORITY = rulib);
(ID = 1)
Name
Kerry Lutz
Role
Copyright holder
Affiliation
Rutgers University. Graduate School - New Brunswick
RightsEvent
(AUTHORITY = rulib);
(ID = 1)
Type
Permission or license
Detail
Non-exclusive ETD license
AssociatedObject
(AUTHORITY = rulib);
(ID = 1)
Type
License
Name
Author Agreement License
Detail
I hereby grant to the Rutgers University Libraries and to my school the non-exclusive right to archive, reproduce and distribute my thesis or dissertation, in whole or in part, and/or my abstract, in whole or in part, in and from an electronic format, subject to the release date subsequently stipulated in this submittal form and approved by my school. I represent and stipulate that the thesis or dissertation and its abstract are my original work, that they do not infringe or violate any rights of others, and that I make these grants as the sole owner of the rights to my thesis or dissertation and its abstract. I represent that I have obtained written permissions, when necessary, from the owner(s) of each third party copyrighted matter to be included in my thesis or dissertation and will supply copies of such upon request by my school. I acknowledge that RU ETD and my school will not distribute my thesis or dissertation or its abstract if, in their reasonable judgment, they believe all such rights have not been secured. I acknowledge that I retain ownership rights to the copyright of my work. I also retain the right to use all or part of this thesis or dissertation in future works, such as articles or books.
Back to the top
Technical
Format
(TYPE = mime);
(VERSION = )
application/x-tar
FileSize
(UNIT = bytes)
48692224
Checksum
(METHOD = SHA1)
53886ae9f4783c4b90b622e3a5a8c45bbe3207f2
ContentModel
ETD
CompressionScheme
other
OperatingSystem
(VERSION = 5.1)
windows xp
Format
(TYPE = mime);
(VERSION = NULL)
application/x-tar
Back to the top
Statistical Profile