TABLE 1.
Plasmids used in this study
| Plasmid | Origina | Relevant genotypeb | Reference or source |
|---|---|---|---|
| pMOSBlue | lacZ bla | Amersham Life Science | |
| pUC7,erm | bla erm | Received from W. de Vos | |
| pCI372 | cat | 12 | |
| pGhost8 | tet orits | 22 | |
| pAK80 | lacLM erm | 15 | |
| pNZ273 | gusA cat | Received from W. de Vos | |
| pTRKH2 | erm | 27 | |
| pCP15 | CP15-lacLM | 16 | |
| pCP22 | CP22-lacLM | 16 | |
| pJM334 | Pupp-lacLM | 23 | |
| pAB201 | pGEM-3Zf(+)::1.4-kb PCR BamHI-SalI | orf1 bla | 6 |
| pLB45 | pGEM-7Zf(+)::2.2-kb EcoRI-XbaI TP901-1 1.1-kb erm | orf1 attP bla erm | 9 |
| pLB61 | pAB201 EcoRI-ClaI 3.4-kb::1.6-kb EcoRI-ClaI pLB45 | orf1 bla | This study |
| pLB65 | pCI372::1.9-kb EcoRI-SalI pLB61 | orf1 cat | This study |
| pLB85 | pBF17a::1.8-kb PstI-HindIII pNZ273 | attP127–333gusA bla erm | This study |
| pLB86 | pBF17b::4.0-kb HindIII-SalI pAK80 | attP127–333lacLM bla erm | This study |
| pLB87 | pBF17b::4.1-kb HindIII-SalI pCP22 | attP127–333 CP22-lacLM | This study |
| pLB88 | pBF17b::4.1-kb HindIII-SalI pCP15 | attP127–333 CP15-lacLM | This study |
| pLB89 | pBF17b::4.9-kb HindIII-SalI pJM334 | attP127–333 Pupp-lacLM | This study |
| pLB95 | pGhost8::1.9-kb EcoRI-SalI pLB61 | orf1 tet orits | This study |
| pBF12 | pMOSBlue::333-bp PCR attP 1.1-kb EcoRI erm | attP1–133bla erm | This study |
| pBF17a | pBF12 SmaI-XbaI Klenow 3.9-kb::237-bp HincII attP from pBF12 | attP127–333bla erm | This study |
| pBF17b | pBF12 SmaI-XbaI Klenow 3.9-kb::237-bp HincII attP from pBF12 | attP127–333bla erm | This study |
| pBF18 | pBF12 SmaI-EcoRV 4.1-kb | attP173–333bla erm | This study |
| pBF20 | pBF12 NdeI-KpnI 3.9-kb::107-bp AseI-KpnI attP from pBF18 | attP173–274bla erm | This study |
| pBF26 | pBF12 SmaI-NdeI 3.9-kb::97-bp AseI blunt PCR (PB1-PB4) | attP186–274bla erm | This study |
| pBF27 | pBF12 SmaI-NdeI 3.9-kb::67-bp AseI blunt PCR (T7-PB5) | attP216–274bla erm | This study |
| pBF30 | pBF12 SmaI-SphI 3.9-kb::56-bp SphI blunt PCR (PB6-PB4) | attP186–241bla erm | This study |
a
Kilobases and base pairs indicate sizes of fragments. The restriction endonuclease enzymes used for digestion are indicated. Ligation is shown by ::. For blunt PCR a polymerase that leaves blunt ends is used. Primers for PCR are indicated in parentheses.
b
The subscript numbers for the attP fragments refer to the base numbers in Fig. 1. orits, temperature-sensitive origin.