'fieldset', '#title' => t('Enter Query Sequence'), '#description' => t('Enter one or more queries in the top text box or use the browse button to upload a file from your local disk. The file may contain a single sequence or a list of sequences. In both cases, the data must be in FASTA format. More information.. '), '#collapsible' => TRUE, '#collapsed' => FALSE, ); $form['pBLAST']['FASTA'] = array( '#type' => 'textarea', '#title' => t('Enter FASTA sequence(s)'), '#description'=>t('Enter query sequence(s) in the text area.'), ); // Upload a file as an alternative to enter a query sequence $form['#attributes']['enctype'] = 'multipart/form-data'; $form['pBLAST']['UPLOAD'] = array( '#prefix' => 'Or upload your query files: ', '#type' => 'file', '#description' => t('The file should be a plain-text FASTA file and not a .doc, .docx, etc. It cannot be greater than 10 Mb in size.'), ); //DATABASE// $form['DB'] = array( '#type' => 'fieldset', '#title' => t('Choose Search Set'), '#description' => t('Choose from one of the protein BLAST databases listed below. You can also use the browse button to upload a file from your local disk. The file may contain a single sequence or a list of sequences. '), '#collapsible' => TRUE, '#collapsed' => FALSE, ); $options = DB_options(); $form['DB']['SELECT_DB'] = array( '#type' => 'select', '#title' => t('Protein BLAST Databases:'), '#options' => $options, '#default_value' => t('Select a database'), ); // Upload a file as an alternative to enter a query sequence $form['#attributes']['enctype'] = 'multipart/form-data'; $form['DB']['DBUPLOAD'] = array( '#prefix' => 'Or upload your own dataset: ', '#type' => 'file', '#description' => t('The file should be a plain-text FASTA file and not a .doc, .docx, etc. It cannot be greater than 10 Mb in size.'), ); //Algorithm parameters $form['ALG'] = array( '#type' => 'fieldset', '#title' => t('Algorithm parameters'), '#collapsible' => TRUE, '#collapsed' => TRUE, ); //General parameters $form['ALG']['GParam'] = array( '#type' => 'fieldset', '#title' => t('General parameters'), '#collapsible' => FALSE, ); $form['ALG']['GParam']['maxTarget'] = array( '#type' => 'select', '#title' => t('Max target sequences:'), '#options' => array( 0 => t('10'), 1 => t('50'), 2 => t('100'), 3 => t('250'), 4 => t('500'), 5 => t('1000'), 6 => t('5000'), 7 => t('10000'), 8 => t('20000'), ), '#default_value' => 2, '#description' => t('Select the maximum number of aligned sequences to display'), ); $form['ALG']['GParam']['shortQueries'] = array( '#type' => 'checkbox', '#title' => t('Automatically adjust parameters for short input sequences'), '#default_value' => TRUE, ); $form['ALG']['GParam']['eVal'] = array( '#type' => 'textfield', '#title' => t('e-value(Expect threshold)'), '#default_value' => 10, '#size' => 12, '#maxlength' => 20, '#description' => t('Expected number of chance matches in a random model.'), ); $form['ALG']['GParam']['wordSize'] = array( '#type' => 'select', '#title' => t('Word size:'), '#options' => array( 0 => t('2'), 1 => t('3'), ), '#default_value' => 1, '#description' => t('The length of the seed that initiates an alignment'), ); $form['ALG']['GParam']['qRange'] = array( '#type' => 'textfield', '#title' => t('Max matches in a query range'), '#default_value' => 0, '#size' => 12, '#maxlength' => 20, '#description' => t('Limit the number of matches to a query range. This option is useful if many strong matches to one part of a query may prevent BLAST from presenting weaker matches to another part of the query.'), ); // Scoring parameters $form['ALG']['SParam'] = array( '#type' => 'fieldset', '#title' => t('Scoring parameters'), '#collapsible' => FALSE, ); $options_first = _ajax_example_get_first_dropdown_options(); $selected = isset($form_state['values']['MATRIX'] ) ? $form_state['values']['MATRIX'] : key($options_first); $form['ALG']['SParam']['MATRIX'] = array( '#type' => 'select', '#title' => 'Matrix', '#options' => $options_first, '#default_value' => $selected, '#description' => t('Assigns a score for aligning pairs of residues, and determines overall alignment score..'), '#ajax' => array( 'callback' => 'ajax_example_dependent_dropdown_callback', 'wrapper' => 'dropdown-second-replace', ), ); $form['ALG']['SParam']['gapCost'] = array( '#type' => 'select', '#title' => t('Gap Costs:'), '#prefix' => '', '#options' => _ajax_example_get_second_dropdown_options($selected), '#default_value' => 2, '#description' => t('Cost to create and extend a gap in an alignment.'), ); $form['ALG']['SParam']['M&MScores'] = array( '#type' => 'select', '#title' => t('Match/Mismatch Scores:'), '#options' => array( 0 => t('No adjustment'), 1 => t('Composition-based statistics'), 2 => t('Conditional compositional score matrix adjustment'), 3 => t('Universal composition score matrix adjustment '), ), '#default_value' => 2, '#description' => t('Matrix adjustment method to compensate for amino acid composition of sequences'), ); //Submit $form['submit'] = array( '#type' => 'submit', '#default_value' => ' BLAST ', ); return $form; } /** * Form validation handler for blast_protein_form(). * * @see blast_protein_form_validate() */ function blast_protein_form_validate($form, &$form_state) { $fastaSeq = $form_state['input']['FASTA']; if (isset($fastaSeq)) { if(validateFasta($fastaSeq)){ form_set_error('pBLAST', t('Error: Failed to read the Blast query: Wrong format provided for FASTA protein sequence')); } else { $form_state['qFlag'] = 'seqQuery'; } } $upQuery = file_save_upload('UPLOAD', array('file_validate_extensions' => array('txt fasta fa fna')), FILE_EXISTS_RENAME); if ($upQuery) { $upQuery_uri = $upQuery->uri; $form_state['upQuery_path'] = drupal_realpath($upQuery_uri); $upQuery_content = file_get_contents($form_state['upQuery_path']); if(validateFasta($upQuery_content)){ form_set_error('pBLAST', t('Error: Failed to upload the Blast query: Wrong format provided for FASTA protein sequence')); } else { $form_state['qFlag'] = 'upQuery'; } } $upDB = file_save_upload('DBUPLOAD', array('file_validate_extensions' => array('txt fasta fa fna')), FILE_EXISTS_RENAME); if ($upDB) { $upDB_uri = $upDB->uri; $form_state['upDB_path'] = drupal_realpath($upDB_uri); $upDB_content = file_get_contents($form_state['upDB_path']); if(validateFasta($upDB_content)){ form_set_error('DB', t('Error: Failed to upload the Blast subject sequence file: Wrong format provided for FASTA protein sequence')); } else { $form_state['dbFlag'] = 'upQuery'; } } else { $form_state['dbFlag'] = 'blastdb'; } } /** * Form submition handler for blast_protein_form(). * * @see blast_protein_form_submit() */ function blast_protein_form_submit($form, &$form_state) { $eVal = $form_state['values']['eVal']; $trgtKey = $form_state['values']['maxTarget']; $numAlign = $form['ALG']['GParam']['maxTarget']['#options'][$trgtKey]; $wsKey = $form_state['values']['wordSize']; $wordSize = $form['ALG']['GParam']['wordSize']['#options'][$wsKey]; $gapKey = $form_state['values']['MATRIX']; switch ($gapKey) { case 0: $matrix ="PAM30"; $gapKey = $form_state['values']['gapCost']; switch ($gapKey) { case 0: $gapOpen = 7; $gapExtend = 2; break; case 1: $gapOpen = 6; $gapExtend = 2; break; case 2: $gapOpen = 5; $gapExtend = 2; break; case 3: $gapOpen = 10; $gapExtend = 1; break; case 4: $gapOpen = 9; $gapExtend = 1; break; case 5: $gapOpen = 8; $gapExtend = 1; break; } break; case 1: $matrix ="PAM70"; $gapKey = $form_state['values']['gapCost']; switch ($gapKey) { case 0: $gapOpen = 8; $gapExtend = 2; break; case 1: $gapOpen = 7; $gapExtend = 2; break; case 2: $gapOpen = 6; $gapExtend = 2; break; case 3: $gapOpen = 11; $gapExtend = 1; break; case 4: $gapOpen = 10; $gapExtend = 1; break; case 5: $gapOpen = 9; $gapExtend = 1; break; } break; case 2: $matrix ="PAM250"; $gapKey = $form_state['values']['gapCost']; switch ($gapKey) { case 0: $gapOpen = 15; $gapExtend = 3; break; case 1: $gapOpen = 14; $gapExtend = 3; break; case 2: $gapOpen = 13; $gapExtend = 3; break; case 3: $gapOpen = 12; $gapExtend = 3; break; case 4: $gapOpen = 11; $gapExtend = 3; break; case 5: $gapOpen = 17; $gapExtend = 2; break; case 6: $gapOpen = 16; $gapExtend = 2; break; case 7: $gapOpen = 15; $gapExtend = 2; break; case 8: $gapOpen = 14; $gapExtend = 2; break; case 9: $gapOpen = 13; $gapExtend = 2; break; case 10: $gapOpen = 21; $gapExtend = 1; break; case 11: $gapOpen = 20; $gapExtend = 1; break; case 12: $gapOpen = 19; $gapExtend = 1; break; case 13: $gapOpen = 18; $gapExtend = 1; break; case 14: $gapOpen = 17; $gapExtend = 1; break; } break; case 3: $matrix ="BLOSUM80"; $gapKey = $form_state['values']['gapCost']; switch ($gapKey) { case 0: $gapOpen = 8; $gapExtend = 2; break; case 1: $gapOpen = 7; $gapExtend = 2; break; case 2: $gapOpen = 6; $gapExtend = 2; break; case 3: $gapOpen = 11; $gapExtend = 1; break; case 4: $gapOpen = 10; $gapExtend = 1; break; case 5: $gapOpen = 9; $gapExtend = 1; break; } break; case 4: $matrix ="BLOSUM62"; $gapKey = $form_state['values']['gapCost']; switch ($gapKey) { case 0: $gapOpen = 11; $gapExtend = 2; break; case 1: $gapOpen = 10; $gapExtend = 2; break; case 2: $gapOpen = 9; $gapExtend = 2; break; case 3: $gapOpen = 8; $gapExtend = 2; break; case 4: $gapOpen = 7; $gapExtend = 2; break; case 5: $gapOpen = 6; $gapExtend = 2; break; case 6: $gapOpen = 13; $gapExtend = 1; break; case 7: $gapOpen = 12; $gapExtend = 1; break; case 8: $gapOpen = 11; $gapExtend = 1; break; case 9: $gapOpen = 10; $gapExtend = 1; break; case 10: $gapOpen = 9; $gapExtend = 1; break; } break; case 5: $matrix ="BLOSUM45"; $gapKey = $form_state['values']['gapCost']; switch ($gapKey) { case 0: $gapOpen = 13; $gapExtend = 3; break; case 1: $gapOpen = 12; $gapExtend = 3; break; case 2: $gapOpen = 11; $gapExtend = 3; break; case 3: $gapOpen = 10; $gapExtend = 3; break; case 4: $gapOpen = 15; $gapExtend = 2; break; case 5: $gapOpen = 14; $gapExtend = 2; break; case 6: $gapOpen = 13; $gapExtend = 2; break; case 7: $gapOpen = 12; $gapExtend = 2; break; case 8: $gapOpen = 19; $gapExtend = 1; break; case 9: $gapOpen = 18; $gapExtend = 1; break; case 10: $gapOpen = 17; $gapExtend = 1; break; case 11: $gapOpen = 16; $gapExtend = 1; break; } break; case 6: $matrix ="BLOSUM50"; $gapKey = $form_state['values']['gapCost']; switch ($gapKey) { case 0: $gapOpen = 13; $gapExtend = 3; break; case 1: $gapOpen = 12; $gapExtend = 3; break; case 2: $gapOpen = 11; $gapExtend = 3; break; case 3: $gapOpen = 10; $gapExtend = 3; break; case 4: $gapOpen = 9; $gapExtend = 3; break; case 5: $gapOpen = 16; $gapExtend = 2; break; case 6: $gapOpen = 15; $gapExtend = 2; break; case 7: $gapOpen = 14; $gapExtend = 2; break; case 8: $gapOpen = 13; $gapExtend = 2; break; case 9: $gapOpen = 12; $gapExtend = 2; break; case 10: $gapOpen = 19; $gapExtend = 1; break; case 11: $gapOpen = 18; $gapExtend = 1; break; case 12: $gapOpen = 17; $gapExtend = 1; break; case 13: $gapOpen = 16; $gapExtend = 1; break; case 14: $gapOpen = 15; $gapExtend = 1; break; } break; case 7: $matrix ="BLOSUM90"; $gapKey = $form_state['values']['gapCost']; switch ($gapKey) { case 0: $gapOpen = 9; $gapExtend = 2; break; case 1: $gapOpen = 8; $gapExtend = 2; break; case 2: $gapOpen = 7; $gapExtend = 2; break; case 3: $gapOpen = 6; $gapExtend = 2; break; case 4: $gapOpen = 11; $gapExtend = 1; break; case 5: $gapOpen = 10; $gapExtend = 1; break; case 6: $gapOpen = 9; $gapExtend = 1; break; } break; } if ( isset($form_state['qFlag']) ) { if ( $form_state['qFlag'] == 'seqQuery' ) { $seq_content = $form_state['values']['FASTA']; $query = "/tmp/user__query_file.fasta"; file_put_contents ( $query , $seq_content); } elseif ( $form_state['qFlag'] == 'upQuery' ) { $query = $form_state['upQuery_path']; } } if ($form_state['dbFlag'] == 'upQuery') { $subjectSeq = $form_state['upDB_path']; $subSeqOut = drupal_basename($form_state['upDB_path']) . rand(0, 10000); $blast_subj_cmd = "blastp -task blastp -query $query -subject $subjectSeq -out sites/default/files/$subSeqOut.blastp.html"; // -evalue $eVal -word_size $wordSize -gapopen $gapOpen -gapextend $gapExtend -matrix $matrix -num_alignments 100 -html"; system($blast_subj_cmd); } elseif ($form_state['dbFlag'] == 'blastdb') { $selected_db = $form_state['values']['SELECT_DB']; $blastdb_node = node_load($selected_db); $blastdb_path = $blastdb_node->db_path; $blastdb_human_name = $form['DB']['SELECT_DB']['#options'][$selected_db]; $subSeqOut = str_replace(' ','_', $blastdb_human_name) . rand(0, 10000); $blast_db_cmd = "blastp -task blastp -query $query -db $blastdb_path -out sites/default/files/$subSeqOut.blastp.html -evalue $eVal -word_size $wordSize -gapopen $gapOpen -gapextend $gapExtend -matrix $matrix -num_alignments 100 -html"; system($blast_db_cmd,$input); } $path = "$subSeqOut.blastp.html"; drupal_goto("blast/report/$path"); } /** * FASTA validating parser * * @param $sequence * A string of characters to be validated. A sequence in FASTA format begins with a single-line description, followed by lines of sequence data. * The description line is distinguished from the sequence data by a greater-than (">") symbol in the first column. * The word following the ">" symbol is the identifier of the sequence, and the rest of the line is the description (both are optional). * There should be no space between the ">" and the first letter of the identifier. The sequence ends if another line starting with a ">" appears; * this indicates the start of another sequence. * * @return * Return a boolean. 1 if the sequence does not pass the format valifation stage and 0 otherwise. * */ function validateFasta($sequence) { $fastaIdRegEx = '/^>.*(\\n|\\r)/'; $fastaSeqRegEx = '/[^acgturykmswbdhvnxACGTURYKMSWBDHVNX\*\-\n\r]/'; if ( preg_match($fastaSeqRegEx,$sequence) && !(preg_match($fastaIdRegEx,$sequence)) ) { $flag = 1; } else { $flag = 0; } return $flag; } /** * Load the preexisting blast databases * * @return * Return human readble names of the preexisting blast databases * */ function DB_options() { $type = 'blastdb'; $nodes = node_load_multiple(array(), array('type'=> $type)); $options = array(); foreach ($nodes as $node) { if ( isset($node) && isset($node->db_dbtype) ) { if ( ($node->db_dbtype=='p') ) { $options[$node->nid] = $node->db_name; } } } asort($options); $options[0] = 'Select a Dataset'; return $options; } /** * Fill the first dropdown list with appropriate options * * @return * An array consisting of matrices name for the first dropdown list * */ function _ajax_example_get_first_dropdown_options() { return drupal_map_assoc(array( t('PAM30'), t('PAM70'), t('PAM250'), t('BLOSUM80'), t('BLOSUM62'), t('BLOSUM45'), t('BLOSUM50'), t('BLOSUM90'), )); } /** * Fill the second dropdown list with appropriate options * * @return * An array containing open and extension gap values for the chosen matrix (to fill the second dropdown list) * */ function _ajax_example_get_second_dropdown_options($key = '') { $options = array( t('PAM30') => drupal_map_assoc(array( t('Existence: 7 Extension: 2'), t('Existence: 6 Extension: 2'), t('Existence: 5 Extension: 2'), t('Existence: 10 Extension: 1'), t('Existence: 9 Extension: 1'), t('Existence: 8 Extension: 1'), )), t('PAM70') => drupal_map_assoc(array( t('Existence: 8 Extension: 2'), t('Existence: 7 Extension: 2'), t('Existence: 6 Extension: 2'), t('Existence: 11 Extension: 1'), t('Existence: 10 Extension: 1'), t('Existence: 9 Extension: 1'), )), t('PAM250') => drupal_map_assoc(array( t('Existence: 15 Extension: 3'), t('Existence: 14 Extension: 3'), t('Existence: 13 Extension: 3'), t('Existence: 12 Extension: 3'), t('Existence: 11 Extension: 3'), t('Existence: 17 Extension: 2'), t('Existence: 16 Extension: 2'), t('Existence: 15 Extension: 2'), t('Existence: 14 Extension: 2'), t('Existence: 13 Extension: 2'), t('Existence: 21 Extension: 1'), t('Existence: 20 Extension: 1'), t('Existence: 19 Extension: 1'), t('Existence: 18 Extension: 1'), t('Existence: 17 Extension: 1'), )), t('BLOSUM80') => drupal_map_assoc(array( t('Existence: 8 Extension: 2'), t('Existence: 7 Extension: 2'), t('Existence: 6 Extension: 2'), t('Existence: 11 Extension: 1'), t('Existence: 10 Extension: 1'), t('Existence: 9 Extension: 1'), )), t('BLOSUM62') => drupal_map_assoc(array( t('Existence: 11 Extension: 2'), t('Existence: 10 Extension: 2'), t('Existence: 9 Extension: 2'), t('Existence: 8 Extension: 2'), t('Existence: 7 Extension: 2'), t('Existence: 6 Extension: 2'), t('Existence: 13 Extension: 1'), t('Existence: 12 Extension: 1'), t('Existence: 11 Extension: 1'), t('Existence: 10 Extension: 1'), t('Existence: 9 Extension: 1'), )), t('BLOSUM45') => drupal_map_assoc(array( t('Existence: 13 Extension: 3'), t('Existence: 12 Extension: 3'), t('Existence: 11 Extension: 3'), t('Existence: 10 Extension: 3'), t('Existence: 15 Extension: 2'), t('Existence: 14 Extension: 2'), t('Existence: 13 Extension: 2'), t('Existence: 12 Extension: 2'), t('Existence: 19 Extension: 1'), t('Existence: 18 Extension: 1'), t('Existence: 17 Extension: 1'), t('Existence: 16 Extension: 1'), )), t('BLOSUM50') => drupal_map_assoc(array( t('Existence: 13 Extension: 3'), t('Existence: 12 Extension: 3'), t('Existence: 11 Extension: 3'), t('Existence: 10 Extension: 3'), t('Existence: 9 Extension: 3'), t('Existence: 16 Extension: 2'), t('Existence: 15 Extension: 2'), t('Existence: 14 Extension: 2'), t('Existence: 13 Extension: 2'), t('Existence: 12 Extension: 2'), t('Existence: 19 Extension: 1'), t('Existence: 18 Extension: 1'), t('Existence: 17 Extension: 1'), t('Existence: 16 Extension: 1'), t('Existence: 15 Extension: 1'), )), t('BLOSUM90') => drupal_map_assoc(array( t('Existence: 9 Extension: 2'), t('Existence: 8 Extension: 2'), t('Existence: 7 Extension: 2'), t('Existence: 6 Extension: 2'), t('Existence: 11 Extension: 1'), t('Existence: 10 Extension: 1'), t('Existence: 9 Extension: 1'), )), ); if (isset($options[$key])) { return $options[$key]; } else { return array(); } } /** * Respond to Ajax dropdown call * * @return * * */ function ajax_example_dependent_dropdown_callback($form, $form_state) { return $form['ALG']['SParam']['gapCost']; }