nBlog 2008-01-11

pNV119 is out!

Last year, I mentioned a high-copy-number mutant of pAL5000 and declared the construction of a high-copy-number version of pNV vector too. Introduction of the 3-bp deletion into pNV19 vector by inverted PCR resulted in a considerable increase of the copy-number of the vector in N. farcinica IFM 10152. I named this mutant pNV119 (version 0).

Taking a leaf from Bourn's paper, I checked the increase of the copy-number of pNV119 by an agarose gel electrophoresis of BamHI-digested total DNA from IFM 10152/pNV119. As shown in the photo, pNV119 was detected clearly (arrow head) but pNV19 was not. This result indicates the increased copy-number of pNV119 if chromosomal and plasmid DNA are extracted with similar efficiencies. Furthermore, pNV119-carrying strain is able to grow in the presence of 1,000 μg/ml of neomycin but pNV19-carrying strain is able to grow in the presence of only up to 500 μg/ml of neomycin (data not shown). This phenotypic difference would also reflect the difference of copy-number between pNV119 and pNV19.

Anyway, I am sure that a high-copy-number version of pNV vector, pNV119, will facilitate our and of course your work. I will prepare pNV119 DNA for distribution as soon as possible.