Table 2.
Plasmids/Constructs
| Construct | Description | Markera | Reference |
|---|---|---|---|
| pCDL280 | pREP1 with GFP (S65T) | AmpR | Varadarajan et al. (2005) |
| pBNB168 | pBlueScript KS II-Ura4 | AmpR | Chen et al. (2004) |
| pBNB189 | ΔG1 (AA276-283): Nucleotides encoding AA1-275 and AA284-470 were generated by PCR using pBNB190 as template with NB380+NB379 and NB378+NB377 sets of respective primers. The two fragments were used in fusion PCR using primers NB380 and NB377. The PCR product was inserted into pCDL280 as in pBNB190. | Amp | This study |
| pBNB190 | The full-length Grn1 gene without its intron was amplified by PCR using S. pombe genomic DNA as template and primers NB380 and NB377. The resulting Grn1 was digested with SalI and NotI and inserted immediately upstream of the GFP gene in vector pCDL280. | AmpR | This study |
| pBNB202 | ΔCC (AA70-90): Nucleotides encoding AA1-69 and AA91-470 were generated by PCR using BNB190 as template with NB380+NB454 and NB453+NB377 sets of respective primers. The resulting fragments were used in fusion PCR with NB380 and NB377 as the primers. The PCR product was inserted into pCDL280 as in pBNB190. | AmpR | This study |
| pBNB203 | ΔG5 (AA164-175): Nucleotides corresponding to AA1-163 and AA176-470 were amplified by PCR using BNB190 as template with NB380+NB458 and NB457-NB377 sets of respective primers. The products were used for fusion PCR with primers NB380 and NB377. The resulting fragment was inserted in pCDL280 as in pBNB190. | AmpR | This study |
| pBNB204 | ΔRG (AA405-415): Nucleotides corresponding to AA1-404 and AA416-470 were amplified by PCR using BNB190 as template with NB380+NB456 and NB455+NB377 sets of respective primers. The PCR products were used for fusion PCR with primers NB380 and NB377. The resulting product was inserted in pCDL280 as in pBNB190. | AmpR | This study |
| pBNB217 | ΔG4 (AA195-208): Nucleotides corresponding to AA1-194 and AA209-470 were amplified by PCR using BNB190 as template with NB380+NB525 and NB524+NB377 sets of respective primers. The PCR products were used for fusion PCR with primers NB380 and NB377. The resulting product was inserted in pCDL280 as in pBNB190. | AmpR | This study |
| pBNB221 | The Rpl25a gene (SPBC106.18) was generated without its intron. The N- and C-terminal fragments were amplified by PCR using the genomic DNA as template with NB535+NB541 and NB540+NB536 sets of respective primers. The fusion product was obtained by PCR using N- and C-terminal products as template with NB535 and NB536 as primers. The resulting product was inserted into pCDL280 using SalI and NotI. | AmpR | This study |
| pBNB284 | The full-length human NGP-1 gene was amplified by PCR using a HeLa cDNA library as template, with primers NB712 and NB713. The PCR product was digested with SalI and NotI and inserted in pCDL280 similarly as in pBNB190. | AmpR | This study |
| pBNB316 | The full-length GNL3L gene was amplified by PCR using a HeLa cDNA library as template and primers NB762 and NB763. The PCR product was cloned into pCDL280 immediately upstream of GFP gene with SalI and NotI. | AmpR | This study |
| pCDNA3.1 | AmpR | Novagen | |
| pBNB335 | ΔG5-GFP fusion was amplified using pBNB203 as template and NB864+NB865 as primers. The PCR was cloned into pCDNA3.1 with KpnI and XhoI. | AmpR | This study |
| pBNB336 | ΔG4-GFP fusion gene was cloned into pCDNA3.1 following the same strategy as in pBNB335 except using pBNB217 as template for PCR. | AmpR | This study |
| pBNB337 | ΔG1-GFP fusion gene was cloned in pCDNA3.1 following the same strategy as in pBNB335 using pBNB189 as template. | AmpR | This study |
| pBNB338 | The Grn1-GFP fusion gene was cloned into pCDNA3.1 following the same strategy as in pBNB335 using pBNB190. | AmpR | This study |
| pBNB339 | ΔRG-GFP fusion gene was cloned into pCDNA3.1 following the same strategy as in pBNB335 using pBNB204. | AmpR | This study |
| pBNB340 | GFP was released from pBNB8 by BamHI/XhoI and inserted in pCDNA3.1 containing the same unique sites. | AmpR | This study |
| pBNB341 | GNL3L-GFP fusion gene was cloned into pCDNA3.1 following the same strategy as pBNB335 cloning using pBNB316 as template and NB883+NB865 as primers. | AmpR | This study |
| pBNB343 | pBlueScript KS II-KanMX6 | AmpR | Chen et al. (2004) |
| pBNB373 | Nucleotides encoding the FLAG epitope were annealed and cloned into pBNB341 with NotI and XhoI. FLAG replaces GFP in this vector. | AmpR | This study |
| pBNB376 | Primers NB907+NB908 were annealed to form a siRNA fragment specific for GNL3L at nt 1047-1065 and inserted in the pSIREN shuttle vector (BD Biosciences). | KanR | This study |
| pBNB377 | Primers NB909+NB910 were annealed to form a scrambled version of GNL3L siRNA (pBNB376) and inserted in pSIREN shuttle vector. | KanR | This study |
| pBNB378 | Oligonucleotides specific for the Luciferase gene were annealed to form a siRNA fragment and inserted in pSIREN shuttle vector. | KanR | This study |
| pBNB395 | Nucleotides encoding the FLAG epitope were annealed and cloned into pBNB316 with NotI and XhoI. FLAG replaces GFP in this vector. | AmpR | This study |
| pBNB396 | The KanMX6 cassette (BNB343) was inserted into pBNB373 using ApaI and XbaI. | AmpR | This study |
| pBNB412 | ΔG3 (AA326-329): Nucleotides encoding AA1-325 and AA330-470 were generated by PCR using BNB190 as template with NB380+NB1007 and NB1006+NB377 sets of respective primers. The two fragments were used in fusion PCR using primers NB380 and NB377. The resulting product was inserted in pCDL280 as in pBNB190. | AmpR | This study |
| pBNB417 | ΔG3-GFP fusion was cloned into pCDNA3.1 following the same strategy as in pBNB335 using pBNB412. | AmpR | This study |
| pBNB494 | The human nucleostemin (NS) gene was amplified by PCR using a HeLa cDNA library as template, with primers NB1160 and NB1161. The PCR product was cloned into pCDL280 following the same strategy as in pBNB190. | AmpR | This study |
| pBNB561 | The S.cerevisiae NUG1 gene was amplified by PCR using yeast genomic DNA as template, with primers NB1309 and NB1310. The PCR product was cloned into pCDL280 following the same strategy as in pBNB190. | AmpR | This study |
a
R, resistance.