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// =============================================================== //
// //
// File : arb_primer.cxx //
// Purpose : //
// //
// Institute of Microbiology (Technical University Munich) //
// http://www.arb-home.de/ //
// //
// =============================================================== //
#include <arbdbt.h>
#include <arb_strarray.h>
#define ADD_LEN 10
#define PRM_BUFFERSIZE 256
struct arb_prm_struct : virtual Noncopyable {
ConstStrArray alignment_names;
int al_len;
int max_name;
GBDATA *gb_main;
char buffer[PRM_BUFFERSIZE];
const char *source;
int prmanz;
int prmlen;
int prmsmin;
char **data;
int sp_count;
int key_cnt;
int one_key_cnt;
int reduce;
FILE *out;
char *outname;
arb_prm_struct()
: al_len(0),
max_name(0),
gb_main(NULL),
source(NULL),
prmanz(0),
prmlen(0),
prmsmin(0),
data(NULL),
sp_count(0),
key_cnt(0),
one_key_cnt(0),
reduce(0),
out(NULL),
outname(NULL)
{
memset(buffer, 0, sizeof(buffer));
}
~arb_prm_struct() {
if (data) {
for (int i = 0; i<sp_count; ++i) free(data[i]);
free(data);
}
free(outname);
}
};
static arb_prm_struct aprm;
inline int getNumFromStdin() {
// returns entered number (or 0)
int i = 0;
if (fgets(aprm.buffer, PRM_BUFFERSIZE, stdin)) {
i = atoi(aprm.buffer);
}
return i;
}
static GB_ERROR arb_prm_menu() {
printf(" Please select an Alignment:\n");
int i;
for (i=1; aprm.alignment_names[i-1]; ++i) {
printf("%i: %s\n", i, aprm.alignment_names[i-1]);
}
aprm.max_name = i;
GB_ERROR error = NULL;
i = getNumFromStdin();
if ((i<1) || (i>=aprm.max_name)) {
error = GBS_global_string("selection %i out of range", i);
}
else {
aprm.source = aprm.alignment_names[i-1];
printf("This module will search for primers for all positions.\n"
" The best result is one primer for all (marked) taxa , the worst case\n"
" are n primers for n taxa.\n"
" Please specify the maximum number of primers:\n"
);
aprm.prmanz = getNumFromStdin();
printf("Select minimum length of a primer, the maximum will be (minimum + %i)\n", ADD_LEN);
i = getNumFromStdin();
if ((i<4) || (i>30)) {
error = GBS_global_string("selection %i out of range", i);
}
else {
aprm.prmlen = i;
printf("There may be short sequences or/and deletes in full sequences\n"
" So a primer normally does not match all sequences\n"
" Specify minimum percentage of species (0-100 %%):\n");
i = getNumFromStdin();
if ((i<1) || (i>100)) {
error = GBS_global_string("selection %i out of range", i);
}
else {
aprm.prmsmin = i;
printf("Write output to file (enter \"\" to write to screen)\n");
if (fgets(aprm.buffer, PRM_BUFFERSIZE, stdin)) {
char *lf = strchr(aprm.buffer, '\n');
if (lf) lf[0] = 0; // remove linefeed from filename
aprm.outname = strdup(aprm.buffer);
}
else {
aprm.outname = strdup("");
}
}
}
}
return error;
}
static GB_ERROR arb_prm_read(int /* prmanz */) {
GBDATA *gb_presets = GBT_get_presets(aprm.gb_main);
{
GBDATA *gb_source = GB_find_string(gb_presets, "alignment_name", aprm.source, GB_IGNORE_CASE, SEARCH_GRANDCHILD);
GBDATA *gb_len = GB_brother(gb_source, "alignment_len");
aprm.al_len = GB_read_int(gb_len);
}
int sp_count = GBT_count_marked_species(aprm.gb_main);
aprm.data = (char **)calloc(sp_count, sizeof(char *));
sp_count = 0;
for (GBDATA *gb_species = GBT_first_marked_species(aprm.gb_main);
gb_species;
gb_species = GBT_next_marked_species(gb_species))
{
GBDATA *gb_source = GB_entry(gb_species, aprm.source);
if (gb_source) {
GBDATA *gb_source_data = GB_entry(gb_source, "data");
if (gb_source_data) {
const char *hdata = GB_read_char_pntr(gb_source_data);
if (!hdata) {
GB_print_error();
}
else {
char *data = (char *)calloc(sizeof(char), aprm.al_len+1);
aprm.data[sp_count ++] = data;
if (sp_count % 50 == 0) printf("Reading taxa %i\n", sp_count);
int size = GB_read_string_count(gb_source_data);
int i;
for (i=0; i<size; i++) {
char c = hdata[i];
if ((c>='a') && (c<='z')) {
data[i] = c-'a'+'A';
}
else {
data[i] = c;
}
}
for (; i<aprm.al_len; i++) {
data[i] = '.';
}
data[i] = 0;
}
}
}
}
printf("%i taxa read\n", sp_count);
aprm.sp_count = sp_count;
if (sp_count == 0) {
return "No marked taxa found";
}
return NULL;
}
static long arb_count_keys(const char * /* key */, long val, void *)
{
if (val >1) {
aprm.key_cnt++;
}
else {
aprm.one_key_cnt++;
}
return val;
}
static long arb_print_primer(const char *key, long val, void *)
{
if (val <= 1) return val;
int gc = 0;
const char *p;
for (p = key; *p; p++) {
if (*p == 'G' || *p == 'C') gc++;
}
fprintf(aprm.out, " %s matching %4li taxa GC = %3i%%\n",
key, val, 100*gc/(int)strlen(key));
return val;
}
#define is_base(c) (((c>='a') && (c<='z')) || ((c>='A')&&(c<='Z')))
static int primer_print(char *dest, char * source, int size)
{
char c;
c = *(source++);
if (!is_base(c)) return 1;
while (size) {
while (!is_base(c)) {
c = *(source++);
if (!c) return 1;
}
if (c == 'N' || c == 'n') return 1;
*(dest++) = c;
size--;
if (!c) return 1;
c = 0;
}
*dest = 0;
return 0;
}
static long arb_reduce_primer_len(const char *key, long val, void *cl_hash) {
GB_HASH* hash = (GB_HASH*)cl_hash;
char buffer[256];
int size = strlen(key)-aprm.reduce;
strncpy(buffer, key, size);
buffer[size] = 0;
val += GBS_read_hash(hash, buffer);
GBS_write_hash(hash, buffer, val);
return val;
}
static void arb_prm_primer(int /* prmanz */)
{
GB_HASH *mhash;
int sp;
char *buffer;
int pos;
int prmlen;
int pspecies;
int cutoff_cnt;
int *best_primer_cnt;
int *best_primer_new;
int *best_primer_swap;
prmlen = aprm.prmlen + ADD_LEN + 1;
buffer = (char *) calloc(sizeof(char), prmlen + 1);
best_primer_cnt = (int *)calloc(prmlen+1, sizeof(int));
best_primer_new = (int *)calloc(prmlen+1, sizeof(int));
for (pos = 0; pos < aprm.al_len; pos++) {
prmlen = aprm.prmlen + ADD_LEN;
mhash = GBS_create_hash(1024, GB_MIND_CASE);
pspecies = 0;
if (pos % 50 == 0) printf("Pos. %i (%i)\n", pos, aprm.al_len);
cutoff_cnt = aprm.prmanz+1;
for (sp = 0; sp < aprm.sp_count; sp++) { // build initial hash table
if (!primer_print(buffer, aprm.data[sp] + pos, prmlen)) {
GBS_incr_hash(mhash, buffer);
pspecies++;
}
}
if (pspecies*100 >= aprm.prmsmin * aprm.sp_count) { // reduce primer length
for (; prmlen >= aprm.prmlen; prmlen-=aprm.reduce) {
GB_HASH *hash = GBS_create_hash(aprm.prmanz, GB_MIND_CASE);
aprm.key_cnt = 0;
aprm.one_key_cnt = 0;
GBS_hash_do_loop(mhash, arb_count_keys, NULL);
if ((aprm.key_cnt + aprm.one_key_cnt < cutoff_cnt) &&
// (aprm.key_cnt > aprm.one_key_cnt) &&
(aprm.key_cnt<best_primer_cnt[prmlen+1])) {
fprintf(aprm.out, "%3i primer found len %3i(of %4i taxa) for position %i\n", aprm.key_cnt, prmlen, pspecies, pos);
GBS_hash_do_loop(mhash, arb_print_primer, NULL);
fprintf(aprm.out, "\n\n");
cutoff_cnt = aprm.key_cnt;
}
best_primer_new[prmlen] = aprm.key_cnt;
aprm.reduce = 1;
while (aprm.key_cnt > aprm.prmanz*4) {
aprm.key_cnt/=4;
aprm.reduce++;
}
GBS_hash_do_loop(mhash, arb_reduce_primer_len, hash);
GBS_free_hash(mhash);
mhash = hash;
}
}
else {
for (; prmlen>0; prmlen--) best_primer_new[prmlen] = aprm.prmanz+1;
}
GBS_free_hash(mhash);
best_primer_swap = best_primer_new;
best_primer_new = best_primer_cnt;
best_primer_cnt = best_primer_swap;
mhash = 0;
}
free(best_primer_new);
free(best_primer_cnt);
free(buffer);
}
int ARB_main(int argc, char *argv[]) {
const char *path = NULL;
while (argc >= 2) {
if (strcmp(argv[1], "--help") == 0) {
fprintf(stderr,
"Usage: arb_primer [dbname]\n"
"Searches sequencing primers\n");
return EXIT_FAILURE;
}
path = argv[1];
argv++; argc--;
}
if (!path) path = ":";
GB_ERROR error = NULL;
GB_shell shell;
aprm.gb_main = GB_open(path, "r");
if (!aprm.gb_main) {
error = GBS_global_string("Can't open db '%s' (Reason: %s)", path, GB_await_error());
}
else {
GB_begin_transaction(aprm.gb_main);
GBT_get_alignment_names(aprm.alignment_names, aprm.gb_main);
GB_commit_transaction(aprm.gb_main);
error = arb_prm_menu();
if (!error) {
GB_begin_transaction(aprm.gb_main);
error = arb_prm_read(aprm.prmanz);
if (!error) {
GB_commit_transaction(aprm.gb_main);
if (strlen(aprm.outname)) {
aprm.out = fopen(aprm.outname, "w");
if (!aprm.out) {
error = GB_IO_error("writing", aprm.outname);
}
else {
arb_prm_primer(aprm.prmanz);
fclose(aprm.out);
}
}
else {
aprm.out = stdout;
arb_prm_primer(aprm.prmanz);
}
}
else {
GB_abort_transaction(aprm.gb_main);
}
}
GB_close(aprm.gb_main);
}
if (error) {
fprintf(stderr, "Error in arb_primer: %s\n", error);
return EXIT_FAILURE;
}
return EXIT_SUCCESS;
}
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