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©The Author(s) 2003.
World J Gastroenterol. Mar 15, 2003; 9(3): 578-583
Published online Mar 15, 2003. doi: 10.3748/wjg.v9.i3.578
Published online Mar 15, 2003. doi: 10.3748/wjg.v9.i3.578
Table 1 Oligonucleotide primers in multiplex PCR of the hu-man HPRT locus
| Exons | Primers sequence (5’-3’) | Fragment size (bp) |
| 1 | F TGG GAC GTC TGG TCC AAG GAT TCA | 626 |
| R CCG AAC CCG GGA AAC TGG CCG CCC | ||
| 2 | F CCT GAT ATG CTC TCA TTG AAA CA | 211 |
| R GCT GCT GAT GTT TGA AAT TAA CAC | ||
| 3 | F GTT TAA TGA CTA AGA GGT GTT TG | 311 |
| R GAA AAC CTA GTG TTG CCA CAT AA | ||
| 4 | F GTG TGT GTA CAT AAG GAT ATA CA | 165 |
| R TTC TTC CCT TTC AAG ATA CAT AC | ||
| 5 | F GGA AAT ACC GTT TTA TTC ATT GT | 125 |
| R GTG CAT ACT AAG TTA GAA AGG TC | ||
| 6 | F GTG ACT CTG AAT TTA AAG CTA TG | 150 |
| R CTG TGT CAA AAT GTC ATA CAT AC | ||
| 7/8 | F GTC TCT CTG TAT GTT ATA TGT CAC | 379 |
| R TGC GTG TTT TGA AAA ATG AGT GAG | ||
| 9 | F GCT ATT CTT GCC TTT CAT TTC AG | 136 |
| R CAA ACT CAA CTT GAA CTC TCA TC |
Table 2 Cytotoxicity and mutagenicity of γ-rays and ENU in HL-60 cells
Table 3 Summary of multiplex PCR analysis of THH-induced HPRT mutants in HL-60 cells
| Categories of mutation | Number analyzed | Number showing PCR changes | Percentage deleted | Number showing no change | |
| Complete deletion | Partial deletion | ||||
| Spontaneous | 13 | 0 | 1/13 (7.7%) | 7.7% | 12/13 (92.3%) |
| ENU (μg/mL) | |||||
| 12.5 | 8 | 1/8 (12.5%) | 4/8 (50.0%)a | 62.5%b | 3/8 (37.5%)a |
| 25.0 | 10 | 2/10 (20.0%) | 5/10 (50.0%)a | 70.0%b | 3/10 (30.0%)b |
| 50.0 | 10 | 2/10 (20.0%) | 6/10 (60.0%)b | 80.0%b | 2/10 (20.0%)b |
| 100.0 | 12 | 4/12 (33.3%)a | 6/12 (50.0%)a | 83.3%b | 2/12 (16.7%)b |
| 200.0 | 19 | 7/19(36.8%)a | 10/19 (52.6%)a | 89.4%b | 2/19 (10.6%)b |
| γ-rays (Gy) | |||||
| 0.5 | 7 | 1/7 (14.3%) | 1/7 (14.3%) | 28.6% | 5/7 (71.4%) |
| 1.0 | 13 | 3/13 (23.1%) | 5/13 (38.5%) | 61.5%b | 5/13 (38.5%)b |
| 2.0 | 10 | 3/10 (30.0%)a | 3/10 (30.0%) | 60.0%b | 4/10 (40.0%)b |
| 4.0 | 17 | 7/17 (41.2%)b | 6/17(35.3%) | 76.5%b | 4/17 (23.5%)b |
Table 4 Schematic diagram of the distribution of deletion within nine exons of the human HPRT gene
| Types of exons deletion | Mutation clones | |||||||||
| 1 | 2 | 3 | 4 | 5 | 6 | 7/8 | 9 | Number | Percent (%) | |
| Spontaneous mutants | 12 | 92.3 | ||||||||
| —a | 1 | 7.7 | ||||||||
| ENU-induced mutants | 12 | 20.3 | ||||||||
| — | 3 | 5.1 | ||||||||
| — | 2 | 3.4 | ||||||||
| — | — | 4 | 6.8 | |||||||
| — | 2 | 3.4 | ||||||||
| — | — | 1 | 1.7 | |||||||
| — | — | — | 2 | 3.4 | ||||||
| — | — | — | 2 | 3.4 | ||||||
| — | — | — | — | 2 | 3.4 | |||||
| — | — | — | 1 | 1.7 | ||||||
| — | — | — | — | 1 | 1.7 | |||||
| — | — | — | — | — | 3 | 5.1 | ||||
| — | — | — | — | — | — | — | 1 | 1.7 | ||
| — | — | — | — | — | — | — | — | 16 | 27.1 | |
| — | — | — | — | — | 1 | 1.7 | ||||
| — | — | — | — | 1 | 1.7 | |||||
| — | — | 2 | 3.4 | |||||||
| — | — | — | — | 2 | 3.4 | |||||
| — | — | 1 | 1.7 | |||||||
| r-rays-induced mutants | 18 | 38.3 | ||||||||
| — | 1 | 2.1 | ||||||||
| — | — | 1 | 2.1 | |||||||
| — | — | 2 | 4.3 | |||||||
| — | — | — | 2 | 4.3 | ||||||
| — | — | — | 1 | 2.1 | ||||||
| — | — | — | — | 1 | 2.1 | |||||
| — | — | — | — | 3 | 6.4 | |||||
| — | — | — | — | — | 1 | 2.1 | ||||
| — | — | — | — | — | 1 | 2.1 | ||||
| — | — | — | — | — | — | — | 2 | 4.3 | ||
| — | — | — | — | — | — | — | — | 14 | 29.8 | |
- Citation: Liu SX, Cao J, An H, Shun HM, Yang LJ, Liu Y. Analysis of spontaneous, gamma ray- and ethylnitrosourea-induced hprt mutants in HL-60 cells with multiplex PCR. World J Gastroenterol 2003; 9(3): 578-583
- URL: https://www.wjgnet.com/1007-9327/full/v9/i3/578.htm
- DOI: https://dx.doi.org/10.3748/wjg.v9.i3.578
