Do you have the gene sequence? (I'm sure you thought of thism, but) I'd start with a literature search or annotate and blast the orfs against public databases. You should get at least a rough idea of function and associated protiens. I'd also look for information about the known pathway(s) where this gene is found.

good luck

Francis-Hoasiuhu said:

I would like to clone a gene of a eukaryotic organism into E.coli DH5λ .Then after I will transfer the cloned plasmid into Origami- for protein expression. I am trying to find out the function and the shape of the protein that the gene is coding for. Can any one help me to make a comment on the above? Note: the protein of interest is soluble thus e-coli is suitable vector since I have only 4 months for the research. I am also wondering if the specific gene/protein is expressive (phenotypic) without the associate protein. Is it possible to clone two gene of interest at the same time/ I am just wondering of the capacity of the plasmid. No idea. I will appreciate any comments. Thanks in advance.

It is possible to clone two genes of interest at the same time if you have a second ribozyme binding site for the second gene. People always use this for expressing two genes (esp. they can form some complex) in the same system. But if you want to find out the functions of each individual protein, then you need to purify them seperately.

thanks very much for the replies i have now epressed my protein fused with 6 histag.i am trying to purify the protein from the 6 histag after using entorokynese.Can any one help me find the easiest way to purify the enzyme and 6 histag.My protein is only 10kD.iam doing ion chromotography for this.any alternatives.