How to find catalytic domain using cDNA
Posted 22 April 2009 - 07:28 AM
got back to this forum after a long time and forgot my old account's password, damn, i'll try to recover it later.
Now i need urgent tips about how to find the catalytic domain of the gene im working on? I need to make its constitutive active form by keeping the catalytic domain only. If anybody could help, ill be really thankful........im also trying to read some literature....
Posted 22 April 2009 - 07:11 PM
Posted 22 April 2009 - 09:28 PM
I believe you first need to find out the ORF in your cDNA. Without knowing whether the DNA sequence encodes protein, there is no meaning of finding catelytic domain. Once you have the ORF, translate it into protein sequence, and then you can use the protein sequence to find domain. There are many tools that allow you to run such analysis such as the InterProScan http://www.ebi.ac.uk...s/InterProScan/
Thanks pcrman, i forgot to mention about the ORF. After translating ORF, i pasted the sequence in the site mentioned by you and found three distinct domains and some unintegrated data. Hence im interested in the catalytic domain only that is responsible for the interaction of my protein. Now what i need to do is to find the sequence coding for the so called catalytic domain. But i tried to track back from the tyr.protein kinase domain (believed to be catalytic) but i did'nt get the sequence, damn i ought to pay attention on bioinfo (thanks to my bioinformatics tachers who did'nt want to teach in english) Plz guide me to find the sequence coding for the desired domain....
Thanks in advance, equal to the number of DNA copies after 500 pcr cycles amplification. ..