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tutorials:eccb_t2_badasp [2012/09/07 15:37] romainstuder |
tutorials:eccb_t2_badasp [2012/09/08 16:15] (current) romainstuder |
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==== BADASP ==== | ==== BADASP ==== | ||
- | BADASP can produce different kind of measure: | + | BADASP can produce different measures: |
* bad: similar the **Type II** of functional divergence. The threshold to choose depend if we want to be stringeant (i.e. BAD > 4) or more relaxed (BAD > 2). | * bad: similar the **Type II** of functional divergence. The threshold to choose depend if we want to be stringeant (i.e. BAD > 4) or more relaxed (BAD > 2). | ||
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</code> | </code> | ||
- | === Execution === | + | === Analysis of the V-type proton ATPase 116 kDa subunit a gene family === |
We want to identify the residues making differences between the **isoforms 1** and **isoforms 4** of the V-type proton ATPase 116 kDa subunit a. | We want to identify the residues making differences between the **isoforms 1** and **isoforms 4** of the V-type proton ATPase 116 kDa subunit a. | ||
+ | First, visualise briefly the multiple alignment in Jalview. (File "badasp_eg.fas" in the badasp folder. | ||
- | <code> | ||
- | cd ./badasp # Folder of installation | ||
- | </code> | ||
Execute **badasp** by importing the multiple alignment in FASTA format ("badasp_eg.fas") and activating the interactive mode (i=1): | Execute **badasp** by importing the multiple alignment in FASTA format ("badasp_eg.fas") and activating the interactive mode (i=1): | ||
- | <code>python badasp.py seqin=badasp_eg.fas i=1</code> | + | |
+ | <code> | ||
+ | cd ./badasp # Folder of installation | ||
+ | python badasp.py seqin=badasp_eg.fas i=1</code> | ||
Badasp will ask for the associated tree, in newick format ("badasp_eg.nsf"): | Badasp will ask for the associated tree, in newick format ("badasp_eg.nsf"): | ||
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=> Press enter | => Press enter | ||
+ | </code> | ||
Display Tree, with two groups of sequences: | Display Tree, with two groups of sequences: | ||
V-type proton ATPase 116 kDa subunit a | V-type proton ATPase 116 kDa subunit a | ||
- | - VPP1 = VPP Isoform 1 (8 genes) | + | * VPP1 = VPP Isoform 1 (8 genes) |
- | - NVL = VPP Isoform 4 (3 genes) | + | * NVL = VPP Isoform 4 (3 genes) |
+ | <code> | ||
Rooted Tree (1000 bootstraps). Branch Lengths given. 21 nodes. <ENTER> to continue. | Rooted Tree (1000 bootstraps). Branch Lengths given. 21 nodes. <ENTER> to continue. | ||
=> Press enter | => Press enter | ||
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</code> | </code> | ||
- | We have a tree and we need to define the two groups to analyse: | + | The tree is now loaded and we need to define the two groups to analyse: |
<code> | <code> | ||
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=> Press enter | => Press enter | ||
- | # We need to split the tree on the node 21, so we need to define two groups from the children nodes 20 (= VPP1 subfamily) and 19 (= VPP4 subfamily) . | + | # We need to split the tree on the node 21, |
+ | # so we need to define two groups from the children nodes 20 (= VPP1 subfamily) and 19 (= VPP4 subfamily) . | ||
=> Press M, then enter. # Manual grouping | => Press M, then enter. # Manual grouping | ||
(Tree displayed) | (Tree displayed) | ||
- | Choice? [default=Q]: c # We collapse node | + | Choice? [default=Q]: c # We collapse nodes |
Node [default=0]: 20 | Node [default=0]: 20 | ||
=> Type VPP1, then Press enter | => Type VPP1, then Press enter | ||
- | Choice? [default=Q]: c # We collapse node | + | Choice? [default=Q]: c # We collapse nodes |
Node [default=0]: 19 | Node [default=0]: 19 | ||
=> Type VPP4, then Press enter | => Type VPP4, then Press enter | ||
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</code> | </code> | ||
- | Badasp will now perform some computation. It will reconstruct the ancestral sequences at each node of the tree, using the [[http:dx.doi.org/10.1186/1471-2105-5-123|GASP (Gapped Ancestral Sequence Prediction) method]]: | + | Badasp will now perform some computations. It will reconstruct the ancestral sequences at each node of the tree, using GASP (ref: http:dx.doi.org/10.1186/1471-2105-5-123 ) |
+ | <code> | ||
Making Ancestral Sequences - Variable PAM Weighting | Making Ancestral Sequences - Variable PAM Weighting | ||
Reading PAM1 matrix from jones.pam | Reading PAM1 matrix from jones.pam |