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Table 1 Thirteen DHFR* insertion site clones and their response to methotrexate

From: Genome position and gene amplification

Name Chr. Chr. band Sequence1 (bp) Exp.2 IC-503 (nM) MTX4 (nM) Expression5 (%) No. of colonies6 Colonies screened7 DHFR* amplicon8 DHFR* gain9
1M-34 9 q34.3 137,580,684 43 120 4,172 26.5 (8.50) 10 4 0
1M-39 13 q32.1 96,707,451 22 65 3,835 0.5 (1.00) 0 0 0
1M-42 8 q22.3 102,162,871 21 75 2,660 5.5 (3.25) 15 13 2
1M-43 11 q13.1 65,526,684 22 65 1,828 13 (10.50) 9 0 3
1M-45 11 q23.3 118,293,388 70 200 5,329 2.5 (3.00) 8 1 6
1M-57 3 q27.1 185,207,363 15 75 2,047 3.5 (3.75) 12 3 8
1M-67 1 p34.3 39,533,783 31 90 2,306 0.5 (1.00) 3 1 0
1M-72 2 q35 217,190,120 16 65 5,077 1.0 (2.00) 2 1 1
1M-73 12 p13.2 12,767,229 20 65 3,116 0.5 (1.00) 1 0 0
1M-75 22 q12.2 28,977,607 32 90 3,307 25.5 (12.75) 3 1 1
1M-83 19 q13.2 45,618,743 14 65 756 1.0 (1.75) 6 0 1
1M-84 17 q12 34,161,068 23 75 3,934 11.0 (11.00) 4 0 0
1M-89 19 q13.33 53,682,371 25 75 4,288 2.0 (1.00) 9 3 0
  1. 1Position in the human genome sequence (UCSC Genome Browser, May 2004 freeze). 2Exp., Direction of DHFR* integration in the genome; arrow to the right indicates that the DHFR* coding sequence is integrated in the direction from the p-arm to the q-arm of the chromosome. 3Methotrexate concentration that causes 50% inhibition of the cell growth after 6 days. 4Methotrexate (MTX) concentration used for selection of resistant colonies. 5Expression of DHFR* measured indirectly by quantitative RT-PCR (EGFP expression normalized to GUSB expression, 2-(dCt) × 100). 6Median number (and interquartile range in parentheses) of resistant colonies per 10 cm plate after 28 days of methotrexate treatment. 7Number of independent resistant colonies screened by array CGH. 8Resistant colonies with amplification of the DHFR* integration region. 9Resistant colonies with low level copy number gain of the DHFR* integration region. Chr., chromosome.