SMD1163 strain毕赤酵母菌株表达宿主菌 BioVector NTCC质粒载体菌种细胞基因保藏中心

SMD1163 strain毕赤酵母菌株表达宿主菌 BioVector NTCC质粒载体菌种细胞基因保藏中心 Description:
SMD1163 (his4 pep4 prb1), SMD1165 (his4 prb1), or SMD1168 (his4 pep4) are a series of protease-deficient strains that may provide a more suitable environment for expression of certain heterologous proteins.
The PEP4 gene encodes proteinase A, a vacuolar aspartyl protease required for the activation of other vacuolar proteases such as carboxypeptidase Y and proteinase B. Proteinase B, prior to processing and activation by proteinase A, has about half the activity of the processed enzyme.
The PRB1 gene codes for proteinase B. Therefore, pep4 mutants display a substantial decrease or elimination in proteinase A and carboxypeptidase Y activities, and partial reduction in proteinase B activity. In the prb1 mutant, only proteinase B activity is eliminated, while pep4 prb1 double mutants show a substantial reduction or elimination in all three of these protease activities.

Recovery
1. Obtain an YPD agar plate.
2. Using a sterile pipette tip, touch the bacteria growing within the punctured area of the stab culture. (A sterilized wire loop or sterile toothpick can be used in place of a sterile pipette tip.)
3. Run this tip lightly over a section of the plate, as shown in the figure, to create streak #1.
4. Using another sterile pipette tip, pass through streak #1 and spread the bacteria over a second section of the plate, to create streak #2.
5. Using a third sterile pipette tip, pass through streak #2 and spread the bacteria over the last section of the plate, to create streak #3.
6. Grow overnight in a 30 PoP C incubator (unless a different growth temperature is indicated on the plasmid datasheet).
7. In the morning, single colonies should be visible. If the bacterial growth is too dense, re-streak onto a new agar plate to obtain single colonies.

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