Genetic and molecular analyses of UniformMu transposon insertion lines
Donald R McCarty1, Masaharu Suzuki, Charles Hunter
1Horticultural Sciences Department, Plant Molecular and Cellular Biology Program, University of Florida, Gainesville, FL, USA.
Methods in Molecular Biology (Clifton, N.J.)
|August 7, 2013
Summary
The UniformMu transposon population provides a valuable resource for maize functional genomics research. This guide details accessing these stocks and offers protocols for genotyping transposon insertion alleles using PCR.
Area of Science:
- Genetics
- Plant Biology
- Molecular Biology
Background:
- The UniformMu transposon population is a significant public resource for maize reverse genetics and functional genomics.
- Access to UniformMu genetic stocks is facilitated through MaizeGDB.org, managed by the Maize Cooperation Stock Center.
Purpose of the Study:
- To describe methods for accessing the UniformMu transposon resource.
- To provide recommended protocols for genotyping transposon insertion alleles in maize.
Main Methods:
- Utilizing online tools at MaizeGDB.org for accessing UniformMu genetic stocks.
- Developing gene-specific polymerase chain reaction (PCR) assays for genotyping transposon insertion alleles.
Main Results:
- Established protocols for accessing and utilizing the UniformMu transposon population.
- Detailed methods for molecular analysis of UFMu insertion lines.
Conclusions:
- The UniformMu resource is accessible and amenable to genetic and molecular analyses.
- Standardized genotyping protocols facilitate research on transposon insertions in maize genes.
More Related Videos
Related Concept Videos
02:57DNA-only Transposons
DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
The donor site from where the transposon is excised is either degraded or...
01:24Transposons
Transposons, or "jumping genes," are small mobile genetic elements (MGEs) that range from 700 to 40,000 base pairs in length. They are found in all organisms and can move within the same chromosome or transfer to different chromosomes. In some cases, transposons can also jump between different host DNA molecules, such as plasmids or viruses, contributing to genetic variability.Barbara McClintock first discovered these mobile genetic elements in the 1940s while studying maize genetics, and she...
02:13Overview of Transposition and Recombination
Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
03:18Non-LTR Retrotransposons
As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...
03:08LTR Retrotransposons
LTR retrotransposons are class I transposable elements with long terminal repeats flanking an internal coding region. These elements are less abundant in mammals compared to other class I transposable elements. About 8 percent of human genomic DNA comprises LTR retrotransposons. Some of the common examples of LTR retrotransposons are Ty elements in yeast and Copia elements in Drosophila.
The internal coding region of LTR retrotransposons and their mechanism of transposition closely resembles a...
The internal coding region of LTR retrotransposons and their mechanism of transposition closely resembles a...
01:16In-vitro Mutagenesis
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.