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Prototheca zopfii Kruger
Prototheca zopfii Kruger
規(guī)格:
貨期:
編號(hào):B232687
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱 Prototheca zopfii Kruger
商品貨號(hào) B232687
Strain Designations 48-Y
Application
degrades petroleum crude oil
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Isolation
Colgate Creek sediment, Baltimore Harbor, Chesapeake Bay, MD, 1973.
Product Format frozen
Type Strain no
Comments
Degrades petroleum.
Petroleum-degradation
Growth on acetate or n-alkanes
Medium ATCC® Medium 28: Emmons' modification of Sabouraud's agar
Growth Conditions
Temperature: 25.0°C
Duration: axenic
Protocol: ATCCNO: 16522 SPEC: This strain is distributed as a freeze-dried preparation. See the general procedures for opening a freeze-dried vial. Aseptically add 0.5 ml of ice cold medium to containing 12% sucrose to the freeze-dried inner shell vial. Once completely rehydrated, aseptically transfer the material to a 100 mm agar plate of the appropriate medium and evenly distribute the material over the surface of the agar with a spread bar. Subculture 4-6 weeks when incubated at 25C. Subculture 6-12 months when incubated at 18C To subculture, transfer a loopful of material to a fresh plate and spread evenly over the surface.
Subcultivation
Protocol: ATCCNO: 16522 SPEC: This strain is distributed as a freeze-dried preparation. See the general procedures for opening a freeze-dried vial. Aseptically add 0.5 ml of ice cold medium to containing 12% sucrose to the freeze-dried inner shell vial. Once completely rehydrated, aseptically transfer the material to a 100 mm agar plate of the appropriate medium and evenly distribute the material over the surface of the agar with a spread bar. Subculture 4-6 weeks when incubated at 25C. Subculture 6-12 months when incubated at 18C To subculture, transfer a loopful of material to a fresh plate and spread evenly over the surface.
Cryopreservation

1.? Harvest cells from a culture that is at or near peak density.? Add 2-3 ml fresh ATCC medium 28 broth to each plate and wash cells into suspension.

2.? Collect cells by centrifugation at 800 x g for 5 min. Adjust the concentration of cells to 2 x 106 - 2 x 107/ml in fresh medium.

3.? While cells are centrifuging prepare a 10% (v/v) solution of sterile methanol in fresh broth medium.

4. Mix the cell preparation and the 10% methanol solution in equal portions. Thus, the final concentration will be 106 - 107 cells/ml and 5% (v/v) Methanol. The time from mixing of the cell preparation and methanol stock solution to the beginning of the freezing process should be no less than 5 min and no greater than 15 min.

5.? Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).

6.?? Place the vials in a controlled rate freezing unit.? From room temperature cool at -1°C/min to -40°C.? If the freezing unit can compensate for the heat of fusion, maintain rate at??????? -1°C/min through the heat of fusion.? At -40°C plunge into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus.? Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.? (The cooling rate in this apparatus is approximately

????? -1°C/min.) ?

7.? The frozen preparations should be stored in either the vapor or liquid phase of a nitrogen refrigerator. Frozen preparations stored below -130°C are stabile indefinitely. Those stored at temperatures above -130°C are progressively less stabile as the storage temperature is elevated. Vials can be stored between -80 and -70°C for no longer than one week.

8.?? To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.

9.?? Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and transfer to a test tube containing 5 ml of ATCC medium 28 broth or to the surface of an ATCC medium 28 agar plate.

10.????????? Incubate a test tube culture upright at 25°C with the cap screwed on loosely (loosened one-half turn); incubate a plate upright at 25°C. Subculture every 4-6 weeks when incubated at 25C, or every 6-12 months when incubated at 18C.

Name of Depositor JD Walker
Year of Origin 1973
References

Pore RS. Prototheca taxonomy. Mycopathologia 90: 129-139, 1985.

Koenig DW, Waed HB. Prototheca zopfii: Kruger strain UMK-B growth on acetate or N alkanes. Appl. Environ. Microbiol. 45: 333-336, 1983.

Walker JD, et al. Petroleum-degrading achlorophyllous algae Prototheca zopfii. Nature 254: 423-424, 1975.

Ueno R, et al. Optimization of heterotrophic culture conditions for n-alkane utilization and phylogenetic position based on the 18S rDNA sequence of a thermotolerant Prototheca zopfii strain. J. Biosci. Bioeng. 94: 160-165, 2002.

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