Chlorella sp. (ATCC® 50258)

Organism: Chlorella sp.  /  Depositor: J Van Etten

Permits and Restrictions

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Strain Designations NC64A
Application
Biofuel production
Biosafety Level 1
Isolation Not applicable
Product Format frozen
Storage Conditions Frozen: -70°C or colder
Freeze-Dried: 2°C to 8°C
Live Culture: See Protocols Section
Type Strain no
Comments
Virus infection
Growth cycle of a virus, PBCV-1
DNA synthesis following Infection with the Virus PBCV-1
Viruses of symbiotic Chlorella-like algae
Lytic viruses
Assembly site of the virus PBCV-1
Ultrastructure studies of PBCV-1 virus
Comparison of viruses
Structural proteins and lipids of PBCV-1 virus
Characterization of viruses
Characterization of gene encoding the DNA methyltransferase from virus NC_1A
Medium Medium 5: Sporulation agar
Medium 847: Algal proteose agar
Growth Conditions
Temperature: 20°C to 25°C
Culture System: Axenic
Cryopreservation Harvest and Preservation
  1. Harvest cells from a culture that is at or near peak density by centrifugation at 800 x g for 5 min.
  2. 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 medium.
  4. Mix the cell preparation and the 10% methanol in equal portions. Thus, the final concentration will be 106 - 107 cells/mL and 5% (v/v) Methanol. The time from the 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 should not be stored above -55°C.
  8. To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the vial just 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 add to a centrifuge tube containing 5 mL of ATCC medium 5 (or ATCC medium 847) without agar. Centrifuge at 300 x g for 5 min.
  10. Remove most of the supernatant (=methanol, which can inhibit growth) and then resuspend the pellet. Transfer the culture to a 16 x 125 mm screw-capped test tube containing 5 mL of ATCC medium 5 broth (or ATCC medium 847 broth), or to the surface of an ATCC medium 5 agar plate (20 x 100 mm Petri plate containing 20 mL of ATCC medium 5 agar). (Alternately, an ATCC medium 847 agar plate can be used; this is a 20 x 100 mm Petri plate containing 20 mL of ATCC medium 847 agar.)
  11. Incubate the culture at 50-100 µEinsteins/m2/s irradiance at 25°C. Maintain under a 14/10h light-dark photoperiod.
Name of Depositor J Van Etten
References

Robock A. Virus infection of culturable Chlorella-like algae and development of a plaque assay. Science 219: 994-996, 1982.

Van Etten JL, et al. Growth cycle of a virus, PBCV-1, tat infects Chlorella-like algae. Virology 126: 117-125, 1983.

Van Etten JL, et al. DNA synthesis in a Chlorella-like alga following infection with the virus PBCV-1. Virology 134: 443-449, 1984.

Van Etten JL, et al. Viruses of symbiotic Chlorella-like algae isolated from Paramecium bursaria and Hydra viridis. Proc. Natl. Acad. Sci. USA 79: 3867-3871, 1982.

Van Etten JL, et al. Lytic viruses infecting a Chlorella-like alga. Virology 140: 135-143, 1985. PubMed: 2981448

Meints RH, et al. Assembly site of the virus PBCV-1 in a Chlorella-like green alga: ultrastructural studies. Virology 154: 240-245, 1986. PubMed: 3750845

Meints RH, et al. Infection of a Chlorella-like alga with the virus, PBCV-1: ultrastructural studies. Virology 138: 341-346, 1984. PubMed: 6495652

Reisser W, et al. A comparison of viruses infecting two different Chlorella-like green algae. Virology 167: 143-149, 1988. PubMed: 2847410

Skrdla MP, et al. Structural proteins and lipids in a virus, PBCV-1, which replicates in a Chlorella-like alga. Virology 135: 308-315, 1984. PubMed: 6740941

Schuster AM, et al. Characterization of viruses infecting a eukaryotic Chlorella-like green alga. Virology 150: 170-177, 1986. PubMed: 3006334

Narva KE, et al. Molecular cloning and characterization of the gene encoding the DNA methyltransferase, M.CviBIII, from Chlorella virus NC-1A. Nucleic Acids Res. 15: 9807-9823, 1987. PubMed: 3320956

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation