Table II.
Plasmid Construction
| Plasmid | Construction | Reference | ||
|---|---|---|---|---|
| pRS313 Backbone | ||||
| pSW363 | NUP145 locus lacking base pairs 2,191–2,850 (the GLFG region) | Emtage et al., 1997 | ||
| pRS315 Backbone | ||||
| pSW503 | Entire KAP95 locus in BamHI in pRS315 | This study | ||
| pSW509 | Entire kap95-L63A locus in BamHI in pRS315 | This study* | ||
| pSW696 | Entire kap95-ΔNES locus in BamHI in pRS315 | This study* | ||
| pRS316 Backbone | ||||
| pSW131 | Entire NUP116 locus in SalI/Xbal of pRS316 | Wente and Blobel, 1994 | ||
| pSW132 | Entire NUP100 locus is SalI/Xbal of pRS316 | Wente and Blobel, 1994 | ||
| pSW190 | Entire NUP145 locus in BamHI/SalI of pRS316 | Emtage et al., 1997 | ||
| pSW271 | Entire KAP95 locus in BamHI of pRS316 | Iovine et al., 1995 | ||
| pSW461 | Insertion of five tandem IgG-binding domains into NsiI site at base pair 990 of NUP116 locus | This study* | ||
| pRS304 Backbone | ||||
| pSW251 | NUP116 locus lacking base pairs 1,260–2,873 (the GLFG region) | This study | ||
| pRS305 Backbone | ||||
| pSW263 | NUP100 locus lacking base pair 890–2,657 (the GLFG region) | This study | ||
| pSW645 | Insertion of five tandem IgG-binding domains into NsiI site at base pair 1,253 of NUP100 locus | This study | ||
| pRS306 Backbone | ||||
| pSW280 | NUP145 locus lacking base pairs 2,191–2,850 (the GLFG region) | |||
| pCH436 Backbone | ||||
| pSW511 | kap95-L63A coding region in BamHI | This study | ||
| pCH358 Backbone | ||||
| pSW368 | SRP1 coding region in BamHI | Murphy et al., 1995 | ||
| pSW392 | Fragment from base pair 890–2,657 (GLFG domain) of NUP100 locus in BamHI | Murphy and Wente, 1996 | ||
| pSW608 | Fragment from base pair 1,294–2,446 of (FXFG domain) NUP1 locus in BamHI | This study | ||
| pGEX-3X Backbone | ||||
| pSW304 | Fragment from base pair 1,260–2,873 (GLFG domain) of NUP116 locus in BamHI | This study | ||
| pSW568 | Fragment from base pair 3,383–3,641 (encoding amino acids 1–77) of KAP95 locus in BamHI/EcoRI | This study | ||
| pSW570 | Fragment from base pair 3,383–3,641 (encoding amino acids 1–77) of kap95-ΔNES locus in BamHI/EcoRI | This study | ||
| pGEX-2T Backbone | ||||
| pPS826 | GSP1 coding region | Corbett et al., 1995 | ||
| pQE-32 Backbone | ||||
| pSW329 | KAP95 coding region in BamHI | This study | ||
| pSW571 | kap95-L63A coding region in BamHI | This study | ||
Vector backbone references for pRS313, pRS315, pRS316, pRS304, pRS305, and pRS306 (Sikorski and Hieter, 1989); pCH436 for LexABD fusion proteins, and pCH358 for GAL4AD fusion proteins (generous gifts of C. Hardy); pQE-32 (Qiagen); pGEX-3X (Pharmacia Fine Chemicals).
*
See Materials and Methods section for a more detailed description.