I am working on the comparison between healthy algae and the algae effected by epiphytic diatoms. I want to compare their properties so for that i want to extract the protein responsible for photosynthesis and i want to check the effects of epiphytes on hypothetical protein 32 (chloroplast). Kindly guide me in this regard. How to extract this protein? How to compare? How to measure?
This is a tuff one. Protein, DNA, RNA
Well as you know, proteins, DNA, RNA, etc. are not easily extracted due to the strong cell wall of algae. This is probably why most of the previous studies focused on the dry matter content of algae.
Extraction can be eased using enzymes that degrade the cell wall. Using lysing buffer can therefore improve the yield. I suggest you to test some kits and protocol of protein extraction and see what is the best on your model. Example: www.sigmaaldrich.com/catalog/product/sigma/pe0220 or www.sigmaaldrich.com/catalog/product/sigma/pe0230 or P-PER Plant Protein Extraction Reagent.
Then to characterize the protein extract, I would say full separation + proteomic is maybe the best way to go since it goes cheaper now. You may need to find collaborators to do it. Else you could run a sds page and cut the band of your hypothetical protein, if you know it size.
For quantitation, most people rely on fluorescence but in your case you don't have any specific probe to the protein, so I am not sure yet, maybe HPLC would work after you get an extract of the protein of interest?
Hope this helps,
Be sure to analyse at least three or more individuals with/without epibiosis. There is a lot of literature on the effects of epibiosis on marine macroalgae (and on mechanisms algae use to deal with epibiosis - see Da Gama et al. 2014 in Adv Bot Res) but I never saw anything like you are trying to do. Are you going to analyse other parameters? the algae are under lab conditions or in the field?
Chapter The Antifouling Defence Mechanisms of Marine Macroalgae
@Bernardo Antonio Perez Da Gama sir, This work is to determine the effects of epiphytes on Rhodophytes (Specific Macroalgae). Other parameters includes micrometabolites and other macrometabolites extraction and comparison.
- Badges
- Science topic
- Similar topics
- Correction