Heterologous expression of the Phycomyces blakesleeanus phytoene dehydrogenase gene (carB) in Mucor circinelloides.
A phytoene dehydrogenase-deficient mutant of Mucor circinelloides accumulating only phytoene was transformed with the gene encoding the corresponding enzyme (carB gene) of Phycomyces blakesleeanus. Carotenoids derived from phytoene were detected in the transformants showing that the P. blakesleeanus carB gene complements the M. circinelloides carB mutation. These newly formed carotenoids accumulated in low quantities, indicating that functional complementation was poor. carB mRNA molecules correctly transcribed were detected in the transformants, but they represented a small proportion of the total population of carB-derived mRNAs, mostly constituted by truncated transcripts and by transcripts longer than the transcript that is functional in Phycomyces. These results showed that the P. blakesleeanus carB gene was expressed in M. circinelloides and suggested that the poor complementation observed was owing, at least in part, to the lack of specificity in the recognition of the transcription initiation and termination signals of the P. blakesleeanus carB gene by the M. circinelloides transcriptional machinery.
Ruiz-Hidalgo
M J
MJ
Area de Genética, Departamento de Microbiología y Genética, Universidad de Salamanca, Plaza de los Doctores de la Reina, s/n. 37007, Salamanca, Spain.
Eslava
A P
AP
Alvarez
M I
MI
Benito
E P
EP
D002338:Carotenoids / Q000378:metabolism; D018076:DNA, Complementary; D015870:Gene Expression; D005800:Genes, Fungal; D009089:Mucor / Q000201:enzymology / Q000235:genetics*; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism*; D010800:Phycomyces / Q000201:enzymology / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014170:Transformation, Genetic
D018076:DNA, Complementary; D011994:Recombinant Proteins; D002338:Carotenoids; D010088:Oxidoreductases; C022398:phytoene dehydrogenase