A1: In all yeasts, host secetory pathway proteases may adversely affect the quality of a secreted recombinant protein. In K. latics, the offending protease is most often an apartyl protease from the protein family pfam00026. We developed an isogenic set of four aspartyl protease deletion strains as a means to address detrimental proteolysis. Each strain harbors a marker-free deletion of a single K. latics gene encoding a secretory pathway aspartyl protease that was identified by bioinformatic analysis of genomic sequence.
Q2: What K.lactis protease deletion strains does NEB sell?
Each strain is sold in a frozen competent cell format and is ready to be transformed by any pKLAC-series or pKLMF-series vector. Additionally, NEB sells a sampler pack that combines one competent cell transformation reaction for each strain (NEB #C1007S).
Q3: Which protease deletion strain should i use?
A3: It is difficult to predict which secretory pathway protease(s) may adversely affect any given secreted recombinant protein's expression. If you suspect that your secreted recombinant protein is susceptible to proteolysis, you should try expression of your protein in each protease deletion strain and compare the quality and yield of the recombinant protein produced.
Q4: What vectors may be used with protease deletion strains?
A4: Protease deletion strains can be transformed by any pKLAC-series or pKLMF-series vector.
Q5: What is the parent background for the protease deletion strains?
A5: Protease genes were deleted from the wild-type K.latics GG799 background.
