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pKD1 based vector for K. lactis expression

pKD1 K. lactis heterologous Protein expression

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#1 ionchannelbk

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Posted 20 July 2016 - 07:34 AM

Hello all,

 

I am working on heterologous protein expression project using the yeast, K. lactis, as the host. I am looking for stable episomal expression vectors for K. lactis. There are lots of reports about successful K. lactis expression using pKD1 based expression vector. I searched for these vectors, but failed in finding a commercially available one. Is there anyone in the forum who happened to have pKD1 expression experiences and know how and where to get pKD1 based expression vector?

 

Thanks in advance!

 

 



#2 827753

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Posted 28 July 2016 - 06:47 PM

It looks like NEB has a K. lactis shuttle vector for $330: https://www.neb.com/...2-pklac2-vector (sorry, missed that you're after an non-integrative vector).

 

 

Worst case scenario you can have it synthesized.  Based on the published sequence (https://www.addgene....uence_vdb/3318/), IDT can send you five ~1000 bp gBlocks to Gibson assemble for $750.

 

It's somewhat AT-biased though, so there may be extra complexity charges, I don't know.

 

You could also save some money by just getting the K. lactis specific sequences synthesized and cloning them into a standard E. coli vector to generate your own shuttle vector. It looks like you would have to ligate an E. coli origin and selectable marker into this vector anyway to make it a shuttle vector.


Edited by 827753, 28 July 2016 - 09:57 PM.


#3 ionchannelbk

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Posted 02 August 2016 - 12:54 PM

Thank you for your advice, 827753. I am thinking of generating my own shuttle episomal vector too. My concern is it may work, or may not work. Then I may get into the chasing-rabbit-hole situation. If I can get a known working pKD1 based vector, then it will be easier to make a decision whether this vector is good to express my protein or not.







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