产品描述信息
ARP-2853?? Toxoplasma gondii Low-Level Pyrimethamine-Resistance Plasmid (pDHFR-TSc3/M3), Recombinant in Escherichia coli(Plasmid/Vectors)|Toxoplasma gondii|Toxoplasma gondii Low-Level Pyrimethamine-Resistance Plasmid (pDHFR-TSc3/M3), Recombinant in Escherichia coli|-60°C or colder?|DS Roos, EO KendallAcknowledgment for publications should read “The following reagent was provided by the NIH AIDS Reagent Program for distribution by BEI Resources, NIAID, NIH: Toxoplasma gondii Low-Level Pyrimethamine-Resistance Plasmid (pDHFR TSc3/M3), Recombinant in Escherichia coli, ARP 2853, contributed by Dr. David S. Roos."|Quantity limit per order for this item is 1. This item can be ordered twice a year. Orders over this limit will be sent to NIAID for approval before shipment. ARP-2853 is an Escherichia coli (E. coli)
DH5α stock containing a plasmid,
pDHFR-RSc3/M3 (also referred to as pDHFR-RSc3/T83N), which confers low-level
resistance to pyrimethamine. The Toxoplasma gondii dihydrofolate reductase-thymidylate synthase (T.
gondii dhfr-ts) minigene, with the 5’ genomic flanking
sequence fused to cDNA, was cloned into the pKS- polylinker HindIII-EcoRI
sites. An additional EcoRI site
occurs near the dhfr-ts junction.
The vector BamHI site has been filled. The wild-type dhfr-ts coding sequence
contains a Thr83(ACC)-Asn(AAC) mutation that confers low-level pyrimethamine
resistance to transfected parasites.
This mutation is analogous to Ser108-Asn in fansidar-resistant Plasmodium
falciparum. ARP-2853
is suitable for stable transformation of T. gondii. Transformants should be isolated at low
frequency (-4), and will typically contain multiple chromosomal or episomal
transgenes, with a copy number roughly proportional to selection pressure. Linked genes may be co-amplified along with
the selectable marker.Note: This plasmid confers resistance to
pyrimethamine in T. gondii,
rendering traditional pyrimethamine/sulfa therapy ineffective. In the event of accidental infection, alternative
treatment (e.g., clindamycin and sulfa) should be administered immediately. Transgenic parasites harboring pyrimethamine
resistance genes are not to be introduced into animals. Each vial contains approximately 500 ?L of E. coli DH5α with pDHFR-TSc3/M3
plasmid in Luria Bertani (LB) broth supplemented with 10% glycerol.