TABLE 2.
Sequences of primers used for DNA amplification and size and localization of the cloned genes
| Genes | Location and PCR primer sequencesa | Restriction enzyme site | Size of DNA fragment (bp) | Localization of DNA fragment (bp)b | GenBank accession no. | Reference |
|---|---|---|---|---|---|---|
| hsp70 | 5′ end, 5′-CATGccatggcGCGAAAAAAGAAAGTCTAACAAAAT-3′ | NcoI | 1,959 | 151–2109 | M27580 | 13 |
| 3′ end, 5′-CGggatccCTCAGGTTTATCAACAATTTCAACA-3′ | BamHI | |||||
| hsp60 | 5′ end, 5′-CATGccatggTCGCTAAAAACATTAAATACAACGA-3′ | NcoI | 1,635 | 383–2017 | M58027 | 8 |
| 3′ end, 5′-CATGagatctATAGTCCATTCCTGCGCCAGGCATTGC-3′ | BglII | |||||
| pgp3 | 5′ end, 5′-CATGccatggGAAATTCTGGTTTTTATTTGTATAA-3′ | NcoI | 795 | 4054–4848 | J03321 | 12 |
| 3′ end, 5′-CATGagatctAGCGTTTGTTTGAGGTATTACCTCT-3′ | BglII | |||||
| MIP | 5′ end, 5′-CATGtcatgaAGAATATATTAAGTTGGATGCTTAT-3′ | BspHId | 732 | 100–831 | X66126, S47522, X53481 | 26 |
| 3′ end, 5′-CGggatccTTCTGTAACAGATACATTATCGTCG-3′ | BamHI | |||||
| hsp10 | 5′ end, 5′-CATGccatggeCAGATCAAGCAACGACCCTCAAGAT-3′ | NcoI | 309 | 37–345 | M58027 | 8 |
| 3′ end, 5′-CATGagatctTTGCAGAACTGCGATAACTTCGCTC-3′ | BgIII |
a
The lowercase characters correspond to restriction enzyme sites.
b
Nucleotides are numbered according to coordinates of the sequences and refer to their position on the GenBank entries.
c
G will be replaced by A with the QuickChange site-directed mutagenesis kit of Stratagene, La Jolla, Calif.
d
The BspHI restriction site is TCATGA which is able to replace the NcoI (CCATGG), since CATG is conserved.
e
G will be replaced by T with the Quick Change site-directed mutagenesis kit of Stratagene.