Document 0832
 DOCN  M9480832
 TI    Cloning and sequencing of the cDNA encoding Pneumocystis carinii inosine
       monophosphate dehydrogenase (IMPDH).
 DT    9410
 AU    O'Gara MJ; Queener SF; Lee CH; Indiana University School of Medicine,
       Indianapolis.
 SO    Abstr Gen Meet Am Soc Microbiol. 1994;94:590 (abstract no. F-13). Unique
       Identifier : AIDSLINE ASM94/94313113
 AB    Pneumocystis carinii is an opportunistic pathogen that causes pneumonia
       in immunocompromised hosts. Chemotherapy is available for treating P.
       carinii pneumonia, but severe toxicity is a significant problem with the
       compounds currently considered drugs of choice. IMP dehydrogenase
       (IMPDH; EC 1.1.1.205) may be a good alternative target for
       anti-Pneumocystis chemotherapy because IMPDH inhibitors have been used
       successfully to treat leukemia. Our lab found the IMPDH inhibitor,
       mycophenolic acid, to be effective in preventing the growth of P.
       carinii in culture. In order to characterize P. carinii IMPDH and assay
       potential inhibitors in vitro, we cloned part of the IMPDH gene.
       Initially, we amplified a 1.3 kb fragment of P. carinii IMPDH from
       genomic DNA using PCR and two degenerate oligonucleotides based on two
       conserved sequences identified in IMPDH from six different species. The
       DNA sequence of the 1.3 kb product indicated the presence of putative
       introns having the characteristic 5'-GT and AG-3' signal. Using a
       combination of RACE (rapid amplification of cDNA ends) and RT-PCR, we
       isolated and cloned the entire cDNA of the P. carinii IMPDH. The
       nucleotide and amino acid sequences (57-59% and 32-63%, respectively)
       revealed significant homology with published sequences of IMPDH genes.
       We are currently working to express P. carinii IMPDH which will be
       assayed to confirm catalytic activity.
 DE    Amino Acid Sequence  Base Sequence  Cloning, Molecular/*METHODS
       Comparative Study  DNA, Complementary/*ANALYSIS  DNA,
       Fungal/ANALYSIS/GENETICS  Gene Amplification  IMP
       Dehydrogenase/*GENETICS  Pneumocystis carinii/*ENZYMOLOGY/*GENETICS
       Polymerase Chain Reaction/METHODS  Sequence Homology, Amino Acid
       Sequence Homology, Nucleic Acid  MEETING ABSTRACT

       SOURCE: National Library of Medicine.  NOTICE: This material may be
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