Background
Methods
Subjects
PCR
Amplified segment | Primer | Annealing temperature |
---|---|---|
SPG31 Ex 1 | F1: ACCAGCTCACCGCCCAATC R1: GTATTAATAGCCCGAGCCACTGG | 56°C/30 cycles |
SPG31 Ex 2 | F2: AAGATAGAGGTGCCCAAGGATG R2: TGAGAACTGTGATGAAGGAACTCC | 58°C/28 cycles |
SPG31 Ex 3 | F3: GGAAGGATAGGGAGAAGGCTAC R3: CCTGAGGTGAGGTTTGAGGC | 58°C/28 cycles |
SPG31 Ex 4 | F4: TTCAAAAGGGCAGTGTTGCTG R4: ATGACTTGCTGGAGGCAAATGA | 58°C/28 cycles |
SPG31 Ex 5 | F5: GCCAAGCCCAATGCTCAGAG R5: GGTCCTTAGCCTGTTCTGTGTGG | 60°C/28 cycles |
SPG31 Ex 6 | F6: AGCTCCCTCTTGGCCTTTGTC R6: GGACTGGGCCTCTCTCAATGA | 58°C/28 cycles |
SPG31 Ex 7 | F7: GAGATTCAGGGAGACCCCAGC R7: GGAGAGAGAAAAGGCCATGTTTG | 60°C/28 cycles |
DHPLC analyses
Amplified segment | WAVE: oven temperature | time shift |
---|---|---|
SPG31 exon 1 | 65°C/70°C | no |
SPG31 exon 2 | 54,5°C/57°C/58°C | no |
SPG31 exon 3 | 59°C/63°C | 63°C: 2,0 |
SPG31 exon 4 | 56°C/57°C/59°C | 59°C: 1,0 |
SPG31 exon 5 | 58°C/61,5°C/64°C | no |
SPG31 exon 6 | 61°C/62°C/63°C | no |
SPG31 exon 7 | 58,6°C/60,6°C | no |
DNA sequencing
RFLP
In silicoanalysis
Results and Discussion
Nucleotide exchange | Amino acid exchange | |
---|---|---|
SPG31 exon 1 | none | |
SPG31 exon 2 | c.60delG | p.6fsX |
c.105+6T>C | ||
SPG31 exon 3 | c.164C>A | p.Thr55Lys |
SPG31 exon 4 | c.183_184insCT | p.7fsX |
SPG31 exon 5 | c.320T>C | p.Leu107Pro |
c.340_347delAGTTACGA | p.69fsX, | |
c.345C>A | p.Try115X | |
SPG31 exon 6 | c.419_420insG | p.45fsX |
c.478delA | p.62fsX | |
SPG31 exon 7 | c.606+43G>T* |
rs-number/Polymorphism | |
---|---|
SPG31 intron 1 | rs1863059 |
SPG31 intron 2 | rs1863058 |
rs1863056 | |
SPG31 exon 4 | rs2276625 |
SPG31 intron 5 | rs12988844 |
SPG31 intron 6 | c.595-19T>G, G allele: 1,2% |
SPG31 3'UTR | c.606+155T>C, C allele: 1,2% |