bool Aligner::Align(const char* query, const Filter& filter,
Alignment* alignment) const
{
if (!translation_matrix_) return false;
if (reference_length_ == 0) return false;
int query_len = strlen(query);
if (query_len == 0) return false;
int8_t* translated_query = new int8_t[query_len];
TranslateBase(query, query_len, translated_query);
const int8_t score_size = 2;
s_profile* profile = ssw_init(translated_query, query_len, score_matrix_,
score_matrix_size_, score_size);
uint8_t flag = 0;
SetFlag(filter, &flag);
s_align* s_al = ssw_align(profile, translated_reference_, reference_length_,
static_cast<int>(gap_opening_penalty_),
static_cast<int>(gap_extending_penalty_),
flag, filter.score_filter, filter.distance_filter, query_len);
alignment->Clear();
ConvertAlignment(*s_al, query_len, alignment);
alignment->mismatches = CalculateNumberMismatch(&*alignment, translated_reference_, translated_query, query_len);
// Free memory
delete [] translated_query;
align_destroy(s_al);
init_destroy(profile);
return true;
}
int main (int argc, char * const argv[]) {
clock_t start, end;
float cpu_time;
gzFile read_fp, ref_fp;
kseq_t *read_seq, *ref_seq;
int32_t l, m, k, match = 2, mismatch = 2, gap_open = 3, gap_extension = 1, path = 0, reverse = 0, n = 5, sam = 0, protein = 0, header = 0, s1 = 67108864, s2 = 128, filter = 0;
int8_t* mata = (int8_t*)calloc(25, sizeof(int8_t));
const int8_t* mat = mata;
char mat_name[16];
mat_name[0] = '\0';
int8_t* ref_num = (int8_t*)malloc(s1);
int8_t* num = (int8_t*)malloc(s2), *num_rc = 0;
char* read_rc = 0;
static const int8_t mat50[] = {
// A R N D C Q E G H I L K M F P S T W Y V B Z X *
5, -2, -1, -2, -1, -1, -1, 0, -2, -1, -2, -1, -1, -3, -1, 1, 0, -3, -2, 0, -2, -1, -1, -5, // A
-2, 7, -1, -2, -4, 1, 0, -3, 0, -4, -3, 3, -2, -3, -3, -1, -1, -3, -1, -3, -1, 0, -1, -5, // R
-1, -1, 7, 2, -2, 0, 0, 0, 1, -3, -4, 0, -2, -4, -2, 1, 0, -4, -2, -3, 5, 0, -1, -5, // N
-2, -2, 2, 8, -4, 0, 2, -1, -1, -4, -4, -1, -4, -5, -1, 0, -1, -5, -3, -4, 6, 1, -1, -5, // D
-1, -4, -2, -4, 13, -3, -3, -3, -3, -2, -2, -3, -2, -2, -4, -1, -1, -5, -3, -1, -3, -3, -1, -5, // C
-1, 1, 0, 0, -3, 7, 2, -2, 1, -3, -2, 2, 0, -4, -1, 0, -1, -1, -1, -3, 0, 4, -1, -5, // Q
-1, 0, 0, 2, -3, 2, 6, -3, 0, -4, -3, 1, -2, -3, -1, -1, -1, -3, -2, -3, 1, 5, -1, -5, // E
0, -3, 0, -1, -3, -2, -3, 8, -2, -4, -4, -2, -3, -4, -2, 0, -2, -3, -3, -4, -1, -2, -1, -5, // G
-2, 0, 1, -1, -3, 1, 0, -2, 10, -4, -3, 0, -1, -1, -2, -1, -2, -3, 2, -4, 0, 0, -1, -5, // H
-1, -4, -3, -4, -2, -3, -4, -4, -4, 5, 2, -3, 2, 0, -3, -3, -1, -3, -1, 4, -4, -3, -1, -5, // I
-2, -3, -4, -4, -2, -2, -3, -4, -3, 2, 5, -3, 3, 1, -4, -3, -1, -2, -1, 1, -4, -3, -1, -5, // L
-1, 3, 0, -1, -3, 2, 1, -2, 0, -3, -3, 6, -2, -4, -1, 0, -1, -3, -2, -3, 0, 1, -1, -5, // K
-1, -2, -2, -4, -2, 0, -2, -3, -1, 2, 3, -2, 7, 0, -3, -2, -1, -1, 0, 1, -3, -1, -1, -5, // M
-3, -3, -4, -5, -2, -4, -3, -4, -1, 0, 1, -4, 0, 8, -4, -3, -2, 1, 4, -1, -4, -4, -1, -5, // F
-1, -3, -2, -1, -4, -1, -1, -2, -2, -3, -4, -1, -3, -4, 10, -1, -1, -4, -3, -3, -2, -1, -1, -5, // P
1, -1, 1, 0, -1, 0, -1, 0, -1, -3, -3, 0, -2, -3, -1, 5, 2, -4, -2, -2, 0, 0, -1, -5, // S
0, -1, 0, -1, -1, -1, -1, -2, -2, -1, -1, -1, -1, -2, -1, 2, 5, -3, -2, 0, 0, -1, -1, -5, // T
-3, -3, -4, -5, -5, -1, -3, -3, -3, -3, -2, -3, -1, 1, -4, -4, -3, 15, 2, -3, -5, -2, -1, -5, // W
-2, -1, -2, -3, -3, -1, -2, -3, 2, -1, -1, -2, 0, 4, -3, -2, -2, 2, 8, -1, -3, -2, -1, -5, // Y
0, -3, -3, -4, -1, -3, -3, -4, -4, 4, 1, -3, 1, -1, -3, -2, 0, -3, -1, 5, -3, -3, -1, -5, // V
-2, -1, 5, 6, -3, 0, 1, -1, 0, -4, -4, 0, -3, -4, -2, 0, 0, -5, -3, -3, 6, 1, -1, -5, // B
-1, 0, 0, 1, -3, 4, 5, -2, 0, -3, -3, 1, -1, -4, -1, 0, -1, -2, -2, -3, 1, 5, -1, -5, // Z
-1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -1, -5, // X
-5, -5, -5, -5, -5, -5, -5, -5, -5, -5, -5, -5, -5, -5, -5, -5, -5, -5, -5, -5, -5, -5, -5, 1 // *
};
/* This table is used to transform amino acid letters into numbers. */
int8_t aa_table[128] = {
23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23,
23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23,
23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23,
23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23, 23,
23, 0, 20, 4, 3, 6, 13, 7, 8, 9, 23, 11, 10, 12, 2, 23,
14, 5, 1, 15, 16, 23, 19, 17, 22, 18, 21, 23, 23, 23, 23, 23,
23, 0, 20, 4, 3, 6, 13, 7, 8, 9, 23, 11, 10, 12, 2, 23,
14, 5, 1, 15, 16, 23, 19, 17, 22, 18, 21, 23, 23, 23, 23, 23
};
/* This table is used to transform nucleotide letters into numbers. */
int8_t nt_table[128] = {
4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4,
4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4,
4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4,
4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4,
4, 0, 4, 1, 4, 4, 4, 2, 4, 4, 4, 4, 4, 4, 4, 4,
4, 4, 4, 4, 3, 3, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4,
4, 0, 4, 1, 4, 4, 4, 2, 4, 4, 4, 4, 4, 4, 4, 4,
4, 4, 4, 4, 3, 3, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4
};
int8_t* table = nt_table;
// Parse command line.
while ((l = getopt(argc, argv, "m:x:o:e:a:f:pcrsh")) >= 0) {
switch (l) {
case 'm': match = atoi(optarg); break;
case 'x': mismatch = atoi(optarg); break;
case 'o': gap_open = atoi(optarg); break;
case 'e': gap_extension = atoi(optarg); break;
case 'a': strcpy(mat_name, optarg); break;
case 'f': filter = atoi(optarg); break;
case 'p': protein = 1; break;
case 'c': path = 1; break;
case 'r': reverse = 1; break;
case 's': sam = 1; break;
case 'h': header = 1; break;
}
}
if (optind + 2 > argc) {
fprintf(stderr, "\n");
fprintf(stderr, "Usage: ssw_test [options] ... <target.fasta> <query.fasta>(or <query.fastq>)\n");
fprintf(stderr, "Options:\n");
fprintf(stderr, "\t-m N\tN is a positive integer for weight match in genome sequence alignment. [default: 2]\n");
fprintf(stderr, "\t-x N\tN is a positive integer. -N will be used as weight mismatch in genome sequence alignment. [default: 2]\n");
fprintf(stderr, "\t-o N\tN is a positive integer. -N will be used as the weight for the gap opening. [default: 3]\n");
fprintf(stderr, "\t-e N\tN is a positive integer. -N will be used as the weight for the gap extension. [default: 1]\n");
fprintf(stderr, "\t-p\tDo protein sequence alignment. Without this option, the ssw_test will do genome sequence alignment.\n");
fprintf(stderr, "\t-a FILE\tFILE is either the Blosum or Pam weight matrix. [default: Blosum50]\n");
fprintf(stderr, "\t-c\tReturn the alignment path.\n");
fprintf(stderr, "\t-f N\tN is a positive integer. Only output the alignments with the Smith-Waterman score >= N.\n");
fprintf(stderr, "\t-r\tThe best alignment will be picked between the original read alignment and the reverse complement read alignment.\n");
fprintf(stderr, "\t-s\tOutput in SAM format. [default: no header]\n");
fprintf(stderr, "\t-h\tIf -s is used, include header in SAM output.\n\n");
return 1;
}
// initialize scoring matrix for genome sequences
for (l = k = 0; LIKELY(l < 4); ++l) {
for (m = 0; LIKELY(m < 4); ++m) mata[k++] = l == m ? match : -mismatch; /* weight_match : -weight_mismatch */
mata[k++] = 0; // ambiguous base
}
for (m = 0; LIKELY(m < 5); ++m) mata[k++] = 0;
if (protein == 1 && (! strcmp(mat_name, "\0"))) {
n = 24;
table = aa_table;
mat = mat50;
} else if (strcmp(mat_name, "\0")) {
// Parse score matrix.
FILE *f_mat = fopen(mat_name, "r");
char line[128];
mata = (int8_t*)realloc(mata, 1024 * sizeof(int8_t));
k = 0;
m = 0;
while (fgets(line, 128, f_mat)) {
if (line[0] == '*' || (line[0] >= 'A' && line[0] <= 'Z')) {
if (line[0] >= 'A' && line[0] <= 'Z') aa_table[(int)line[0]] = aa_table[(int)line[0] + 32] = m;
char str[4], *s = str;
str[0] = '\0';
l = 1;
while (line[l]) {
if ((line[l] >= '0' && line[l] <= '9') || line[l] == '-') *s++ = line[l];
else if (str[0] != '\0') {
*s = '\0';
mata[k++] = (int8_t)atoi(str);
s = str;
str[0] = '\0';
}
++l;
}
if (str[0] != '\0') {
*s = '\0';
mata[k++] = (int8_t)atoi(str);
s = str;
str[0] = '\0';
}
++m;
}
}
if (k == 0) {
fprintf(stderr, "Problem of reading the weight matrix file.\n");
return 1;
}
fclose(f_mat);
n = m;
table = aa_table;
mat = mata;
}
//fprintf(stderr, "query: %s\n", argv[optind + 1]);
read_fp = gzopen(argv[optind + 1], "r");
if (! read_fp) {
fprintf (stderr, "gzopen of '%s' failed.\n", argv[optind + 1]);
exit (EXIT_FAILURE);
}
read_seq = kseq_init(read_fp);
if (sam && header && path) {
fprintf(stdout, "@HD\tVN:1.4\tSO:queryname\n");
ref_fp = gzopen(argv[optind], "r");
ref_seq = kseq_init(ref_fp);
while ((l = kseq_read(ref_seq)) >= 0) fprintf(stdout, "@SQ\tSN:%s\tLN:%d\n", ref_seq->name.s, (int32_t)ref_seq->seq.l);
kseq_destroy(ref_seq);
gzclose(ref_fp);
} else if (sam && !path) {
fprintf(stderr, "SAM format output is only available together with option -c.\n");
sam = 0;
}
// alignment
if (reverse == 1 && n == 5) {
read_rc = (char*)malloc(s2);
num_rc = (int8_t*)malloc(s2);
}
start = clock();
while (kseq_read(read_seq) >= 0) {
s_profile* p, *p_rc = 0;
int32_t readLen = read_seq->seq.l;
int32_t maskLen = readLen / 2;
while (readLen >= s2) {
++s2;
kroundup32(s2);
num = (int8_t*)realloc(num, s2);
if (reverse == 1 && n == 5) {
read_rc = (char*)realloc(read_rc, s2);
num_rc = (int8_t*)realloc(num_rc, s2);
}
}
for (m = 0; m < readLen; ++m) num[m] = table[(int)read_seq->seq.s[m]];
p = ssw_init(num, readLen, mat, n, 2);
if (reverse == 1 && n == 5) {
reverse_comple(read_seq->seq.s, read_rc);
for (m = 0; m < readLen; ++m) num_rc[m] = table[(int)read_rc[m]];
p_rc = ssw_init(num_rc, readLen, mat, n, 2);
}else if (reverse == 1 && n == 24) {
fprintf (stderr, "Reverse complement alignment is not available for protein sequences. \n");
return 1;
}
ref_fp = gzopen(argv[optind], "r");
ref_seq = kseq_init(ref_fp);
while (kseq_read(ref_seq) >= 0) {
s_align* result, *result_rc = 0;
int32_t refLen = ref_seq->seq.l;
int8_t flag = 0;
while (refLen > s1) {
++s1;
kroundup32(s1);
ref_num = (int8_t*)realloc(ref_num, s1);
}
for (m = 0; m < refLen; ++m) ref_num[m] = table[(int)ref_seq->seq.s[m]];
if (path == 1) flag = 2;
result = ssw_align (p, ref_num, refLen, gap_open, gap_extension, flag, filter, 0, maskLen);
if (reverse == 1 && protein == 0)
result_rc = ssw_align(p_rc, ref_num, refLen, gap_open, gap_extension, flag, filter, 0, maskLen);
if (result_rc && result_rc->score1 > result->score1 && result_rc->score1 >= filter) {
if (sam) ssw_write (result_rc, ref_seq, read_seq, read_rc, table, 1, 1);
else ssw_write (result_rc, ref_seq, read_seq, read_rc, table, 1, 0);
}else if (result && result->score1 >= filter){
if (sam) ssw_write(result, ref_seq, read_seq, read_seq->seq.s, table, 0, 1);
else ssw_write(result, ref_seq, read_seq, read_seq->seq.s, table, 0, 0);
} else if (! result) return 1;
if (result_rc) align_destroy(result_rc);
align_destroy(result);
}
if(p_rc) init_destroy(p_rc);
init_destroy(p);
kseq_destroy(ref_seq);
gzclose(ref_fp);
}
end = clock();
cpu_time = ((float) (end - start)) / CLOCKS_PER_SEC;
fprintf(stderr, "CPU time: %f seconds\n", cpu_time);
if (num_rc) {
free(num_rc);
free(read_rc);
}
kseq_destroy(read_seq);
gzclose(read_fp);
free(num);
free(ref_num);
free(mata);
return 0;
}
