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#ifndef BAMCONVERTER_H_
#define BAMCONVERTER_H_
#include<cstdio>
#include<cstring>
#include<cassert>
#include<string>
#include<map>
#include <stdint.h>
#include "htslib/sam.h"
#include "sam_utils.h"
#include "SamHeader.hpp"
#include "utils.h"
#include "my_assert.h"
#include "bc_aux.h"
#include "Transcript.h"
#include "Transcripts.h"
class BamConverter {
public:
BamConverter(const char* inpF, const char* outF, const char* chr_list, Transcripts& transcripts, int nThreads, const std::string& command);
~BamConverter();
void process();
private:
samFile *in, *out;
bam_hdr_t *in_header, *out_header;
Transcripts& transcripts;
std::map<std::string, int> refmap;
std::map<std::string, int>::iterator iter;
CollapseMap collapseMap;
void convert(bam1_t*, const Transcript&);
void writeCollapsedLines();
void flipSeq(uint8_t*, int);
void flipQual(uint8_t*, int);
void modifyTags(bam1_t*, const Transcript&); // modify MD tag and XS tag if needed
};
BamConverter::BamConverter(const char* inpF, const char* outF, const char* chr_list, Transcripts& transcripts, int nThreads, const std::string& command)
: transcripts(transcripts)
{
general_assert(transcripts.getType() == 0, "Genome information is not provided! RSEM cannot convert the transcript bam file!");
in = sam_open(inpF, "r");
assert(in != 0);
in_header = sam_hdr_read(in);
assert(in_header != 0);
transcripts.buildMappings(in_header->n_targets, in_header->target_name);
SamHeader hdr(in_header->text);
hdr.replaceSQ(chr_list);
hdr.insertPG("rsem-tbam2gbam", command);
// hdr.addComment("This BAM file is processed by rsem-tbam2gam to convert from transcript coordinates into genomic coordinates.");
out_header = hdr.create_header();
refmap.clear();
for (int i = 0; i < out_header->n_targets; ++i) {
refmap[out_header->target_name[i]] = i;
}
out = sam_open(outF, "wb");
assert(out != 0);
sam_hdr_write(out, out_header);
if (nThreads > 1) general_assert(hts_set_threads(out, nThreads) == 0, "Fail to create threads for writing the BAM file!");
}
BamConverter::~BamConverter() {
bam_hdr_destroy(in_header);
sam_close(in);
bam_hdr_destroy(out_header);
sam_close(out);
}
void BamConverter::process() {
bam1_t *b, *b2;
std::string cqname, qname;
bool isPaired = false;
HIT_INT_TYPE cnt = 0;
cqname = "";
b = bam_init1(); b2 = bam_init1();
while (sam_read1(in, in_header, b) >= 0) {
++cnt;
isPaired = bam_is_paired(b);
if (isPaired) {
assert(sam_read1(in, in_header, b2) >= 0 && bam_is_paired(b2));
if (!bam_is_read1(b)) { bam1_t *tmp = b; b = b2; b2 = tmp; }
assert(bam_is_read1(b) && bam_is_read2(b2));
general_assert((bam_is_mapped(b) && bam_is_mapped(b2)) || (bam_is_unmapped(b) && bam_is_unmapped(b2)), \
"Detected partial alignments for read " + bam_get_canonical_name(b) + ", which RSEM currently does not support!");
++cnt;
}
if (cnt % 1000000 == 0) { printf("."); fflush(stdout); }
qname = bam_get_canonical_name(b);
if (bam_is_mapped(b)) {
// for collapsing
if (isPaired) general_assert(b->core.tid == b2->core.tid, qname + "'s two mates are aligned to two different transcripts!");
const Transcript& transcript = transcripts.getTranscriptViaEid(b->core.tid + 1);
convert(b, transcript);
if (isPaired) {
convert(b2, transcript);
b->core.mpos = b2->core.pos;
b2->core.mpos = b->core.pos;
}
if (cqname != qname) {
writeCollapsedLines();
cqname = qname;
collapseMap.init(isPaired);
}
uint8_t *p = bam_aux_get(b, "ZW");
float prb = (p != NULL? bam_aux2f(p) : 1.0);
collapseMap.insert(b, b2, prb);
}
else {
assert(cqname != qname);
writeCollapsedLines();
cqname = qname;
collapseMap.init(isPaired);
sam_write1(out, out_header, b);
if (isPaired) sam_write1(out, out_header, b2);
}
}
writeCollapsedLines();
bam_destroy1(b);
bam_destroy1(b2);
if (cnt >= 1000000) printf("\n");
}
void BamConverter::convert(bam1_t* b, const Transcript& transcript) {
int pos = b->core.pos;
int readlen = b->core.l_qseq;
general_assert(readlen > 0, "One alignment line has SEQ field as *. RSEM does not support this currently!");
iter = refmap.find(transcript.getSeqName());
assert(iter != refmap.end());
b->core.tid = iter->second;
if (bam_is_paired(b)) { b->core.mtid = b->core.tid; }
b->core.qual = 255; // set to not available temporarily
if (transcript.getStrand() == '-') {
b->core.flag ^= BAM_FREVERSE;
if (bam_is_paired(b)) {
b->core.flag ^= BAM_FMREVERSE;
b->core.isize = -b->core.isize;
}
flipSeq(bam_get_seq(b), readlen);
flipQual(bam_get_qual(b), readlen);
}
std::vector<uint32_t> data;
data.clear();
int core_pos, core_n_cigar;
tr2chr(transcript, pos + 1, pos + readlen, core_pos, core_n_cigar, data);
assert(core_pos >= 0);
int rest_len = b->l_data - b->core.l_qname - b->core.n_cigar * 4;
b->l_data = b->core.l_qname + core_n_cigar * 4 + rest_len;
expand_data_size(b);
uint8_t* pt = b->data + b->core.l_qname;
memmove(pt + core_n_cigar * 4, pt + b->core.n_cigar * 4, rest_len);
for (int i = 0; i < core_n_cigar; ++i) { memmove(pt, &data[i], 4); pt += 4; }
b->core.pos = core_pos;
b->core.n_cigar = core_n_cigar;
b->core.bin = hts_reg2bin(b->core.pos, bam_endpos(b), 14, 5);
modifyTags(b, transcript); // check if need to add XS tag, if need, add it
}
inline void BamConverter::writeCollapsedLines() {
bam1_t *tmp_b = NULL,*tmp_b2 = NULL;
float prb;
bool isPaired;
uint8_t *p;
if (!collapseMap.empty(isPaired)) {
while (collapseMap.next(tmp_b, tmp_b2, prb)) {
p = bam_aux_get(tmp_b, "ZW");
if (p != NULL) {
memcpy(bam_aux_get(tmp_b, "ZW") + 1, (uint8_t*)&(prb), bam_aux_type2size('f'));
tmp_b->core.qual = bam_prb_to_mapq(prb);
}
// otherwise, just use the MAPQ score of the orignal alignment
sam_write1(out, out_header, tmp_b);
if (isPaired) {
if (p != NULL) memcpy(bam_aux_get(tmp_b2, "ZW") + 1, (uint8_t*)&(prb), bam_aux_type2size('f'));
tmp_b2->core.qual = tmp_b->core.qual;
sam_write1(out, out_header, tmp_b2);
}
bam_destroy1(tmp_b);
if (isPaired) bam_destroy1(tmp_b2);
}
}
}
inline void BamConverter::flipSeq(uint8_t* s, int readlen) {
uint8_t code, base;
std::vector<uint8_t> seq;
code = 0; base = 0;
seq.clear();
for (int i = 0; i < readlen; ++i) {
switch (bam_seqi(s, readlen - i - 1)) {
case 1: base = 8; break;
case 2: base = 4; break;
case 4: base = 2; break;
case 8: base = 1; break;
case 15: base = 15; break;
default: assert(false);
}
code |= base << (4 * (1 - i % 2));
if (i % 2 == 1) { seq.push_back(code); code = 0; }
}
if (readlen % 2 == 1) { seq.push_back(code); }
for (int i = 0; i < (int)seq.size(); ++i) s[i] = seq[i];
}
inline void BamConverter::flipQual(uint8_t* q, int readlen) {
int32_t mid = readlen / 2;
uint8_t tmp;
for (int i = 0; i < mid; ++i) {
tmp = q[i]; q[i] = q[readlen - i - 1]; q[readlen - i - 1] = tmp;
}
}
inline void BamConverter::modifyTags(bam1_t* b, const Transcript& transcript) {
char strand = transcript.getStrand();
uint8_t *s = NULL;
if (strand == '-') {
s = bam_aux_get(b, "MD");
if ((s != NULL) && (*(s) == 'Z') && (bam_aux2Z(s) != NULL)) {
char *mis = bam_aux2Z(s);
int len = strlen(mis);
char *tmp = new char[len];
int cur_type = -1, fr = -1, type, base;
for (int i = 0; i < len; i++) {
type = (mis[i] >= '0' && mis[i] <= '9');
if (cur_type != type) {
switch(cur_type) {
case 0:
base = len - 1;
if (mis[fr] == '^') { tmp[len - i] = mis[fr]; ++fr; ++base; }
for (int j = fr; j < i; j++) tmp[base - j] = ((mis[j] == 'A' || mis[j] == 'C' || mis[j] == 'G' || mis[j] == 'T') ? getOpp(mis[j]) : mis[j]);
break;
case 1:
base = len - i - fr;
for (int j = fr; j < i; j++) tmp[base + j] = mis[j];
break;
}
cur_type = type;
fr = i;
}
}
switch(cur_type) {
case 0:
base = len - 1;
if (mis[fr] == '^') { tmp[0] = mis[fr]; ++fr; ++base; }
for (int j = fr; j < len; j++) tmp[base - j] = ((mis[j] == 'A' || mis[j] == 'C' || mis[j] == 'G' || mis[j] == 'T') ? getOpp(mis[j]) : mis[j]);
break;
case 1:
for (int j = fr; j < len; j++) tmp[j - fr] = mis[j];
break;
}
strncpy(mis, tmp, len);
delete[] tmp;
}
}
// append XS:A field if necessary
s = bam_aux_get(b, "XS");
if (s != NULL) bam_aux_del(b, s);
bool hasN = false;
uint32_t* p = bam_get_cigar(b);
for (int i = 0; i < (int)b->core.n_cigar; i++)
if ((*(p + i) & BAM_CIGAR_MASK) == BAM_CREF_SKIP) { hasN = true; break; }
if (hasN) bam_aux_append(b, "XS", 'A', 1, (uint8_t*)&strand);
}
#endif /* BAMCONVERTER_H_ */
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