Positive control
@Yujeong Oh
Gene selection
EF1-α
- eukaryotic translation elongation factor 1 alpha
- Function:
- This is used as a housekeeping gene for African clawed frog by several papers Dhorne-Pollet et al, 2013
- suggested to be a housekeeping gene for amphibians by one paper.
- RNA sequence is used in PCR reaction (hence, it requires transcription of RNA to cDNA and downstream amplification)
- Questions when this is used as reference gene for general amphibians/ etc
- When searched this gene on ncbi website, this gene was only appeared on African and tropical clawed frog among amphibians (this gene is present in many other eukaryotes).
- The housekeeping gene are highly conserved, but yet its sequence may not be exactly identical from my understanding. So could this gene of African clawed frog be used as reference gene for other amphibian species (especially when the sequence of other species couldn’t be found)
- Could genomic DNA sequence be used instead of mRNA? Zhang et al., 2016 Dhorne-Pollet et al, 2013
- Tatiersky et al., 2015 uses this gene as reference gene and used the published primers designed for African clawed frog’s EF1-α for amplification while testing the expression level of antimicrobial peptides on leopard frog.
References
| (Author, year) | Link | Title |
|---|---|---|
| Dhorne-Pollet et al, 2013 | https://anatomypubs.onlinelibrary.wiley.com/doi/pdf/10.1002/dvdy.23972 | Validation of Novel Reference Genes for RT-qPCR Studies of Gene Expression in Xenopus Tropicalis During Embryonic and Post-Embryonic Development |
| Zhang et al., 2016 | https://www.nature.com/articles/srep37549 | Selection of reliable reference genes for normalization of quantitative RT-PCR from different developmental stages and tissues in amphioxus |
| Tatiersky et al., 2015 | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4529697/ | Effect of glucocorticoids on expression of cutaneous antimicrobial peptides in northern leopard frogs (Lithobates pipiens) |