Notes

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Analysis of the fugu multidrug resistance gene
  • Decide the starting and stoping sites of MDR1 and MDR3
  • Cut the contig (39951bp) into three parts with some overlaps
  • Fragment 1:         1-14000
  • Fragment 2: 12001-26000
  • Fragment 3: 24001-39951
  • Repeats found with Dotplot analysis
  • Clustalw result of the repeat sequence
  • GCG graphics result of the repeat sequence
  • No 
    Start-end
    Start-end
    Start-end
    Start-end
    Start-end
    Repaet1
    14-49
        51-229
    231-278
    280-334
    336-475
    Repeat2
    12393-12428
    12430-12608
    12610-12657
    12659-12713
    12715-12854
    Repeat3
    23080-23115
    23117-23295
    23297-23344
    23346-23400
    23402-23541
    Length
    36
    179
    48
    55
    140
    Sequence CCATGG-TATACA ACTGTC-TTCACA TAGTGC-TCTTGT TCCAGG-CATCGT GTAGCC-GATGGG
  • Dived the sequence into 3 parts:
  • MDR1: 1-12390
  • MDR3: 12371-23075
  • FRAG3: 23061-39951
  • Align MDR1_HUMAN N and C terminal to determine domains in MDR1
  • MDR1N_HUMAN:  1-666 (666 AA)
  • MDR1C_HUMAN:  667-1280 (614 AA)
  • Retrive MDR1 and MDR3 AA sequence from SWISSPROT

  •  
    Code No AA Oganism 
    MDR1_HUMAN 1280 Human
    MDR1_LEIEN 1280 Leshmania enriettii 
    MDR1_MOUSE 1276 Mouse
    MDR1_RAT 1277 Rat
    MDR1_CRIGR 1276 Chinese hamster 
    MDR1_CAEEL 1321 C. elegans 
    MDR3_HUMAN 1279 Human
    MDR3_MOUSE 1276 Mouse
    MDR3_CRIGR 1281 Chinese Hamster 
    MDR3_CAEEL 1254 C. elegans 
  • Multiple sequence alignment  - WebGene output AA sequence allignment to find best conserved fragments in MDR1 and MDR3 gene
  • Alignment  1 - Based on fragment 1-12390 and 12371-23075
  • Alignment  2 - Based on fragment 1-14000 and 12000-26000 together with 1-12390 and 12371-23075
  • Alignment  3 - Based on Gene_Finder FGenesH for fragment 1-14000 and 12000-26000
  • Alignment  4 - Edited Gene_finder FGenesH result
  • Problems in gene1: the 1st exon of both N and C terminal subunit was missed!

  •  
  • Gene 2 analysis
  • Alignment of gene 2 aa sequence with human MDR1 and MDR 3
  • All 28 exons in gene 2 were icked up!
  • C elegans MDR exons has different structure
  • (NEW!!!)
  • exon 12  was wrong in Gene finder output
  • exon 13 was mixed in exon 12 in Gene Finder output
  • Intron 12 was mannualy picked up with Staden
  • Exon 26  was picked up manually (to be checked)
  • Extra base problem  Email to Yu Jun
  • Gene 2 cDNA sequnece with translation
  • Gene 2 amino acid sequnece
  • Alignment of gene 2 cDNA with human MDR3 DNA
  • Alignment of gene 2 amino acis with human MDR3 DNA
  • Based on FgenH output, we can assign the 2 MDR genes.
  • No    TSS    Exon-S   Exon-E   PolyA  Gene-S  Gene-E  Gene-L
    ----------------------------------------------------------- 
    0                               980            1000 
    1    2279    2667     12870   13091    2279   13105   10827
    2   13221   13741     23557   23696   13221   23710   10490
    
    
  • Gene 1 analysis
  • Some exons are missed in gene 1 prediction
  • exon comparison of human and fugu
  • Local aliangment of gene 1 and gene 2 DNA sequence
  • Global aliangment of gene 1 and gene 2 DNA sequence
  • Gene 2 exon numbering based on the original contig sequence

  •  


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    Last modified: Sat, 21 Aug 1999, office@cbi.pku.edu.cn