Table 1.
Mutation analysis using single-strand conformation polymorphism in RNA and direct DNA sequencing in the reporter genelacI
| SSCP-RNA | Direct DNA sequencing | ||
|---|---|---|---|
| Spontaneous lacI mutants | |||
| Clone no. 50 | nt 548–763 | 569-C to t | (CAG t AAA) |
| 52/54 | nt 50–163 | 86-G to t | (ACC t TTT) |
| 53 | nt 50–163 | 54-T to c | (ATG c CGC) |
| Forebrain ischemia-induced lacI mutants | |||
| 30 min ischemia + 8 hr reperfusion | |||
| Clone no. 10 | nt 723–963 | d813-T | (GGC - GCG) |
| 12 | nt 723–963 | d781-G | (GCT - GGC)* |
| 14 | nt 50–163 | 90-CCC to aag | (TTT aag GCG)* |
| 22 | nt 723–963 | d761-G | (GTT - CCA) |
| d764-AA | (GCC -- CGA) | ||
| d800-A | (ATT - CCG) | ||
| 31 | nt 723–963 | 819-T to a | (GCG a TGG) |
| 35 | nt −49 to 163 | 49-C to a | (ATA a GAT) |
| 39 | nt 723–963 | 843-G to a | (TGG a ATA)* |
| 40 | nt 181–403 | d273-cgattaaatctcgcg-287 | (CGC-n15-CCG) |
| 41 | nt 723–963 | 792-G to a | (TGC a CGC) |
| 42 | nt 723–963 | d927-G | (CCA - CGT) |
| 48 | nt −49 to 163 | 49-C to a | (ATA a GAT) |
| RC-8 | nt −49 to 163 | 92-C to t | (TCC t GCG)* |
| 30 min ischemia + 24 hr reperfusion | |||
| Clone no. 4 | nt 50–251 | 95-G to a | (CGC a TGG) |
| 5 | nt 723–963 | 853-T to g | (CGA g ACC) |
| 6 | nt 50–163 | d50-G | (TAC - ATG) |
| i54-TgC-55 | (TGT g CGC) | ||
| nt 347–601 | 419-C to t | (GAC t AGG) | |
| 7 | nt 548–763 | i675-GgG-676 | (ACG g GGA)* |
| 8 | nt 548–763 | 630-G to a | (GCT a GCA) |
| 9 | nt 548–763 | d624-ctgg-627 | (TGG ---- CTG) |
| 33 | nt 723–963 | 880-G to a | (CCC a CCG) |
Repeatable and identical base alterations were accepted as the mutation of that clone. Upper cases = parental sequences; lower cases = mutant sequences; d = deletion; i = insertions; nt = nucleotide number of lacI gene; * = mutation in the triplet CCC (or GGG) sequences; these mutations were similar to those using nitric oxide (Wink et al., 1991; Nguyen et al., 1992; Marragos et al., 1993;Schmutte et al., 1994).