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Producing unknown protein

About microscopic forms of life, including Bacteria, Archea, protozoans, algae and fungi. Topics relating to viruses, viroids and prions also belong here.

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Producing unknown protein

Postby drfuture » Sat May 18, 2013 8:43 pm

Dear all,


We have protein sequence which is unknown, when we checked any databases we see that it is probably new. the aim is how we can make this protein produced via vectors using molecular approaches? What can be experimental design.
drfuture
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Postby JackBean » Mon May 20, 2013 9:42 am

just clone it into your favour plasmid and express in any host. Do you have any idea what activity it has or how did you get the sequence?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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Postby Cat » Mon May 20, 2013 1:56 pm

Drfuture, your question is too vague. Protein form which organism? Do you have DNA sequence or just protein?

General answer to your question:

1. Backtranslate protein sequence to DNA sequence.
2. Codon optimize to the expression system (you have a choice of bacterial or cell free expression systems)
3. Synthesize sequence (IDT can do it for you).
4. Ligate into vector (vector depends on the expression system)/clone vector.
3. Transform and/or express ...
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