TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or descriptiona | Sources or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| XL1-Blue-MRF′ | Δ(mcrA) 183Δ(mcrCB hsdSMR mrr) 173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac[F′ proAB laqqZΔM15 Tn10] | Gibco BRL |
| DM1 | F−dam−13::Tn9 (Cmr) dcm− mcrB HsdR− M−gal1 gal2 Ara− Lac− Thr− Leu− Tonr Tsxrsup0 | Gibco BRL |
| X. dendrorhous (P. rhodozyma) | ||
| CBS 6938 | Wild-type strain (DNA donor strain) | |
| PR-1-104 | β-Carotene-accumulating mutant of CBS 6938 | 13 |
| Plasmids | ||
| pUC19 | General cloning vector for E. coli (Ap) | 22 |
| pMTL22P | General cloning vector for E. coli (Ap) | 31 |
| pPRGDH6 | 5.5-kb EcoRI fragment carrying the gpd gene of X. dendrorhous in pUC19 (Ap) | 28 |
| pPR1 | General transformation vector for X. dendrorhous | 35 |
| pPR2TN | Derivative of pPR1 with terminator sequences of the gpd gene of X. dendrorhous and additional unique cloning sites (Ap, G418) | 31 |
| pPRcrtYB | cDNA encoding the phytoene synthase-lycopene cyclase of X. dendrorhous in pBluescript (Ap) | 31 |
| pPR10F/R | 4.5-kb EcoRI fragment carrying the genomic copy of the crtYB gene in pUC19 (Ap) | 31 |
| pPR11 | BamHI-NcoI PCR fragment containing Pgpd-crtYB cloned in the 6.3-kb BamHI-NcoI site of pPR10F (Ap) | This study |
| pPR12 | Expression cassette (Pgpd crtYB TcrtYB) as BamHI fragment in pMTL22P (Ap, G418) | This study |
| pPR13F/R | BamHI fragment (Pgpd crtYB TcrtYB) cloned in corresponding site of pPR2TN (Ap, G418) | This study |
| pPR16 | BamHI-HindIII (crtYBΔ) fragment cloned in corresponding sites of pPR1 (G418) | This study |
| pPR19F/R | Insertion of selection cassette (Pgpd G418 Tgpd) on blunt ended (Klenow) EcoRI fragment in the T4 DNA polymerase-treated BstXI site of pPR10F (crtYB::Pgpd G418 Tgpd) (Ap, G418) | This study |
| pPR21 | crtYB gene of X. dendrorhous as a 4.5-kb BamHI fragment in pMTL22P (Ap) | 31 |
| pPR22F/R | 4.5-kb BamHI fragment from pPR21 in BamHI site of pPR2TN (Ap, G418) | 31 |
| pPRpd2 | cDNA encoding phytoene desaturase of X. dendrorhous in pBluescript (Ap) | 30 |
| pPRpdcos1 | Phytoene desaturase-positive cosmid clone from a genomic library of X. dendrorhous (Ap) | 30 |
| pPR40F/R | 2.8-kb BamHI PCR fragment containing the expression cassette for phytoene desaturase (Pcrti crtl Tcrtl) cloned in BamHI site of pPR2TN (Ap, G418) | This study |
a
The selective antibiotics are indicated in parentheses.