[Construction of Cryptococcus neoformans cap70 transformation system].
OBJECTIVE
To construct a transformation system of Cryptococcus neoformans capsule-deficient strain cap70.


METHODS
(1)Obtained ura5 mutants by screening the Cryptococcus neoformans cap70 using 5-fluoroorotic acid counter selection method, analyzed the obtaining strain by Southern blot and PCR; (2)Through the plasmid pCXJ18 and pCXJU which contained ura5 gene from Cryptococcus neoformans and ura3 from Kluyveromyces fragilils. as a selective marker, the ura5 strains were transformed by electrotransformation and chemical transformation.


RESULTS
Obtained ura5 mutants, they were detected as lack of ura function. The chemical transformation method was first be used in Cryptococcus neoformans transformation.


CONCLUSIONS
A transformation system of Cryptococcus neoformans cap70 has been established, which provides not only the basis for gene cloning from the species for studying gene function and gene expression.
Pan
W
W
Department of Dermatology, Changzheng Hospital, The Second military Medical University, Shanghai 200003, China.
Liao
W
W
Huo
K
K
D003455:Cryptococcus neoformans / Q000235:genetics* / Q000502:physiology; D004269:DNA, Bacterial / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D014169:Transformation, Bacterial / Q000235:genetics* / Q000502:physiology
D004269:DNA, Bacterial