Hi, I have tried to verify the function of one enzyme while my target protein is insoluble in E.coli expression system , I have two thoughts:
1. trying the yeast expression system
2. try to find a homologous protein to verify the function indirectly and I have gained the BLAST result from NCBI by PSI-BLAST, they have total 1500 results.
My questions is:
1. I built the phylogenetic tree based on the results BUT now I have no idea about how to analyze the tree and screen the suitable protein I want...Can I just choose the high identity score ones or the nearest protein in phylogenetic tree?
2. my target protein is a glycosyltransferase, is that any specific motif of this kind of enzyme?
3. Can the platform UniProt realize the similar function as BLAST? Based on UniProt's database?
https://www.researchgate.net/post/How_to_interpret_the_results_from_PSI-BLAST
https://www.ncbi.nlm.nih.gov/books/NBK2590/
The above may be helpful to start with your questions. Good luck.
@Shin Murakami really thanks for your recommendation, it made me a further understandind of the basic rules of PSI-BLAST~
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