Cloning in vector with C-terminus epitope

[Shivani Rajoriya]

Hello everyone,

I am trying to clone a gene in a vector with C-terminus epitope. To get a complete protein, I have to add additional nucleotides at 3' end of my gene sequence. Otherwise, a stop codon appears in starting of epitope sequence. 

By doing this, there is change of amino acids between my gene sequence and epitope sequence. Does this change the final protein structure ?

Thank you.

Regards,

Shivani Rajoriya.

[駱子瑜]

Hi, in this case you have no choice but to remove the stop codon of your gene then you can acquire the fusion protein with tag.

And for the additional nucleotides between your gene and tag, it general should not be a problem. However, it is hard to say it will not have any adverse effect. In contrast to the weakness of restriction enzyme based method, other seamless techniques can clone genes into vector without adding extra nucleotides. Search these seamless cloning methods such as HiFiNEBuilder, In fusion cloning …and so on

Good luck

Shivani Rajoriya <shivanii...@gmail.com>於 2023年11月2日 週四，下午9:56寫道：

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