Related Experiment Videos
Murine MLP: cloning and expression in the embryonic head
G V Harrod1, P J Kettunen, A K Jowett
1Department of Biomedical Science, University of Sheffield, United Kingdom.
Abstract:
Differential display reverse transcription polymerase chain reaction (DD RT-PRC) was used to isolate cDNA clones of mRNA expressed in particular compartments of the developing murine head. One clone has been characterized and found to be the murine homologue of the MLP genes. In other species the LIM-domain containing MLPs are expressed specifically in muscle. In situ hybridization and RT-PCR have confirmed that the murine gene is expressed during both cardiac and skeletal myogenesis. Our data show that DD RT-PCR is a valuable technique to clone cDNA clones from minute tissue samples by virtue of the gene's expression pattern rather than by a process of library screening by homology.
Insights
Differential display reverse transcription polymerase chain reaction identified a murine MLP gene crucial for muscle development. This method efficiently clones genes from small tissue samples based on expression patterns.
Area of Science:
- Developmental Biology
- Molecular Biology
- Genetics
Background:
- The murine homologue of LIM-domain containing MLP genes, typically muscle-specific in other species, was investigated.
- Understanding gene expression in specific compartments of the developing murine head is crucial for developmental studies.
Purpose of the Study:
- To isolate and characterize cDNA clones of specific mRNAs from developing murine head compartments.
- To identify novel genes involved in murine myogenesis using differential display techniques.
Main Methods:
- Differential display reverse transcription polymerase chain reaction (DD RT-PCR) was employed to isolate specific mRNA transcripts.
- In situ hybridization and standard RT-PCR were used to confirm gene expression patterns.
Main Results:
- A cDNA clone representing the murine homologue of MLP genes was successfully isolated.
- Expression of this murine MLP gene was confirmed in both cardiac and skeletal myogenesis.
Conclusions:
- DD RT-PCR is an effective method for cloning cDNAs from limited tissue samples by leveraging gene expression profiles.
- The identified murine MLP gene plays a role in both cardiac and skeletal muscle development.