Is it possible to express eukaryotic selenoproteins in E. coli cells.
Yes, It is possible but expression is very little and some time no expression have been observed. I am also trying to increase its expression by adding sodium selenate and/or sodium selenite alog with some chemical regular like sodium butyrate. It has been reported that sodium butyrate some times help in expression and transportation of protein. But, at this stage I am not sure...May be attached review help you at some extent in understanding of selenoprotein expression..Good luck for your research!
Mr. Navin, Thanks for your response.
I am aware that E.coli also having Selenoprotein unlike Yeast and higher plant. Does this SECIS insertion machinery and other Tras acting proteins is suffice to express the heterologous protein (if you knw any good paper describing these or some one knows workign in this field for quit a some time). I do have some theory behind Selenoprotein synthesis. If possible do give your gmail or other ID for regular follow up.
I will read the review which you have give me.
It is interesting review..could you come across any recent paper where in they expressed any eukaryotic selenoprotein in E.coli. Does common JM 109/Dh5alpha will support the selenoprotein expression.
I read one paper where they reported expression of human selnoprotein in Chalamydomonas..model organism for algae (pl. refer the enclosed..).
Dear Dr. A.Muthu,
Thanks for discussion.The topic you are discussing is very least worked up on till date. No one got success, however many approaches have been applied. I am also trying to do the same as a side project but it is try SECIS machinery and other components are not sufficient for good expression level (as I believe). There must be some other important thing which could be extra supplement or any other molecular factors that might help in its good expression level.Although, I am again attaching a paper, may be you know it (not a recent paper) which has nice information...My email id is : [email protected]
Also, Here is weblink of Dr. Sharon Rozovsky reserach group. She has more experience in these fileds. You may approach her for help if she could able to do then.
http://sites.udel.edu/rozovskygroup/home/research/
Yes, expression and purification of eukaryotic selenoprotein but you will need to modify the sequence a little to fit the expression machinery in E. coli cells also you need special plasmid which will provide the E. coli cells with the Se-insertion machinery. an examples for such proteins, prof. Gladyshev (Harvard Medical school) has many articles about MSRB1 and TrxR (both selenoprotein) expression in E. coli.
if still have more questions, I will be happy to assist you to find the answers.
in the attached article, a very good description of how to purify selenoproteins in E. coli.
Thanks for your answers. I will go through the article. Are you working on selenoproteins? If so at what capacity.student or PDF or faculty. Is it possible to get the plasmid construct which is equipped with SECIS sequences? Due credit will be given in publication.
You are welcome, now I am not working on selenoproteins, but as I suggested in my answer and I still recommend you to contact prof. Gladyshev's lab if you want to talk about the plasmid and the credit for using it.
if you need any other information, here is my email [email protected]
Good luck
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