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#include<cstdio>
#include<cstring>
#include<cctype>
#include<cstdlib>
#include<fstream>
#include<sstream>
#include<set>
#include<map>
#include<vector>
#include<algorithm>
#include<string>
#include "utils.h"
#include "my_assert.h"
#include "GTFItem.h"
#include "Transcript.h"
#include "Transcripts.h"
using namespace std;
bool verbose = true;
struct ChrInfo {
string name;
size_t len;
ChrInfo(const string& name, size_t len) {
this->name = name;
this->len = len;
}
bool operator< (const ChrInfo& o) const {
return name < o.name;
}
};
int M;
vector<GTFItem> items;
vector<string> seqs;
vector<int> starts; // used to generate .grp
map<string, vector<int> > sn2tr; // map from seqname to transcripts
map<string, vector<int> >::iterator iter;
vector<ChrInfo> chrvec;
Transcripts transcripts;
char groupF[STRLEN], tiF[STRLEN], refFastaF[STRLEN];
char chromListF[STRLEN];
bool hasMappingFile;
char mappingFile[STRLEN];
map<string, string> mi_table; // mapping info table
map<string, string>::iterator mi_iter; //mapping info table's iterator
set<string> sources;
void parseSources(char* sstr) {
char* p = strtok(sstr, ",");
while (p != NULL) {
sources.insert(p);
p = strtok(NULL, ",");
}
}
inline bool isTrusted(const string& source) {
return sources.size() == 0 || sources.find(source) != sources.end();
}
void loadMappingInfo(char* mappingF) {
ifstream fin(mappingF);
string line, key, value;
general_assert(fin.is_open(), "Cannot open " + cstrtos(mappingF) + "! It may not exist.");
mi_table.clear();
while (getline(fin, line)) {
line = cleanStr(line);
if (line[0] == '#') continue;
istringstream strin(line);
strin>>value>>key;
mi_table[key] = value;
}
fin.close();
}
bool buildTranscript(int sp, int ep) {
int cur_s, cur_e; // current_start, current_end
vector<Interval> vec;
string transcript_id = items[sp].getTranscriptID();
string gene_id = items[sp].getGeneID();
string gene_name = "", transcript_name = "";
char strand = items[sp].getStrand();
string seqname = items[sp].getSeqName();
string left = items[sp].getLeft();
vec.clear();
cur_s = cur_e = -1;
for (int i = sp; i <= ep; i++) {
int start = items[i].getStart();
int end = items[i].getEnd();
general_assert(strand == items[i].getStrand(), "According to the GTF file given, transcript " + transcript_id + " has exons from different orientations!");
general_assert(seqname == items[i].getSeqName(), "According to the GTF file given, transcript " + transcript_id + " has exons on multiple chromosomes!");
if (items[i].getGeneName() != "") {
if (gene_name == "") gene_name = items[i].getGeneName();
else general_assert(gene_name == items[i].getGeneName(), "Transcript " + transcript_id + " is associated with multiple gene names!");
}
if (items[i].getTranscriptName() != "") {
if (transcript_name == "") transcript_name = items[i].getTranscriptName();
else general_assert(transcript_name == items[i].getTranscriptName(), "Transcript " + transcript_id + " is associated with multiple transcript names!");
}
if (cur_e + 1 < start) {
if (cur_s > 0) vec.push_back(Interval(cur_s, cur_e));
cur_s = start;
}
cur_e = (cur_e < end ? end : cur_e);
}
if (cur_s > 0) vec.push_back(Interval(cur_s, cur_e));
transcripts.add(Transcript(transcript_id, gene_id, seqname, strand, vec, left, transcript_name, gene_name));
return true;
}
void parse_gtf_file(char* gtfF) {
ifstream fin(gtfF);
string line, tid, gid;
GTFItem item;
general_assert(fin.is_open(), "Cannot open " + cstrtos(gtfF) + "! It may not exist.");
int cnt = 0;
int n_warns = 0;
items.clear();
while (getline(fin, line)) {
if (line[0] == '#') continue; // if this line is comment, jump it
item.parse(line);
if (item.getFeature() == "exon" && isTrusted(item.getSource())) {
if (item.getStart() > item.getEnd()) {
if (++n_warns <= MAX_WARNS) {
fprintf(stderr, "Warning: exon's start position is larger than its end position! This exon is discarded.\n");
fprintf(stderr, "\t%s\n\n", line.c_str());
}
}
else if (item.getStart() < 1) {
if (++n_warns <= MAX_WARNS) {
fprintf(stderr, "Warning: exon's start position is less than 1! This exon is discarded.\n");
fprintf(stderr, "\t%s\n\n", line.c_str());
}
}
else {
item.parseAttributes(line);
if (hasMappingFile) {
tid = item.getTranscriptID();
mi_iter = mi_table.find(tid);
general_assert(mi_iter != mi_table.end(), "Mapping Info is not correct, cannot find " + tid + "'s gene_id!");
gid = mi_iter->second;
item.setGeneID(gid);
}
items.push_back(item);
}
}
++cnt;
if (verbose && cnt % 200000 == 0) { printf("Parsed %d lines\n", cnt); }
}
fin.close();
if (n_warns > 0) fprintf(stderr, "Warning: In total, %d exons are discarded.", n_warns);
sort(items.begin(), items.end());
int sp = 0, ep; // start pointer, end pointer
int nItems = items.size();
sn2tr.clear();
while (sp < nItems) {
tid = items[sp].getTranscriptID();
ep = sp + 1;
while (ep < nItems && items[ep].getTranscriptID() == tid) ep++;
ep--;
buildTranscript(sp, ep);
int sid = transcripts.getM();
const Transcript& transcript = transcripts.getTranscriptAt(sid);
iter = sn2tr.find(transcript.getSeqName());
if (iter == sn2tr.end()) {
vector<int> vec(1, sid);
sn2tr[transcript.getSeqName()] = vec;
}
else {
iter->second.push_back(sid);
}
sp = ep + 1;
}
items.clear();
M = transcripts.getM();
general_assert(M > 0, "The reference contains no transcripts!");
if (verbose) { printf("Parsing gtf File is done!\n"); }
}
void shrink() {
int curp = 0;
int n_warns = 0;
for (int i = 1; i <= M; i++)
if (seqs[i] == "") {
if (++n_warns <= MAX_WARNS) {
const Transcript& transcript = transcripts.getTranscriptAt(i);
fprintf(stderr, "Warning: Cannot extract transcript %s's sequence since the chromosome it locates, %s, is absent!\n", transcript.getTranscriptID().c_str(), transcript.getSeqName().c_str());
}
}
else {
++curp;
transcripts.move(i, curp);
if (i > curp) seqs[curp] = seqs[i];
}
if (n_warns > 0) fprintf(stderr, "Warning: %d transcripts are failed to extract because their chromosome sequences are absent.\n", n_warns);
if (verbose) printf("%d transcripts are extracted.\n", curp);
transcripts.setM(curp);
M = transcripts.getM();
general_assert(M > 0, "The reference contains no transcripts!");
starts.clear();
string curgid = "", gid;
for (int i = 1; i <= M; i++) {
gid = transcripts.getTranscriptAt(i).getGeneID();
if (curgid != gid) {
starts.push_back(i);
curgid = gid;
}
}
starts.push_back(M + 1);
}
void writeResults(char* refName) {
int s;
ofstream fout;
sprintf(groupF, "%s.grp", refName);
sprintf(tiF, "%s.ti", refName);
sprintf(refFastaF, "%s.transcripts.fa", refName);
sprintf(chromListF, "%s.chrlist", refName);
fout.open(groupF);
s = starts.size();
for (int i = 0; i < s; i++) fout<<starts[i]<<endl;
fout.close();
if (verbose) { printf("Group File is generated!\n"); }
transcripts.writeTo(tiF);
if (verbose) { printf("Transcript Information File is generated!\n"); }
fout.open(chromListF);
s = chrvec.size();
for (int i = 0; i < s; i++) {
fout<<chrvec[i].name<<'\t'<<chrvec[i].len<<endl;
}
fout.close();
if (verbose) { printf("Chromosome List File is generated!\n"); }
fout.open(refFastaF);
for (int i = 1; i <= M; i++) {
fout<<">"<<transcripts.getTranscriptAt(i).getTranscriptID()<<endl;
fout<<seqs[i]<<endl;
}
fout.close();
if (verbose) { printf("Extracted Sequences File is generated!\n"); }
}
struct CursorPos {
char *filename;
int line_no, pos;
} cursor;
inline char check(char c) {
general_assert(isalpha(c), "FASTA file " + cstrtos(cursor.filename) + " contains an unknown character, " + \
ctos(c) + " (ASCII code " + itos(c) + "), at line " + itos(cursor.line_no) + ", position " + itos(cursor.pos + 1) + "!");
if (isupper(c) && c != 'A' && c != 'C' && c != 'G' && c != 'T') c = 'N';
if (islower(c) && c != 'a' && c != 'c' && c != 'g' && c != 't') c = 'n';
return c;
}
int main(int argc, char* argv[]) {
if (argc < 7 || ((hasMappingFile = atoi(argv[5])) && argc < 8)) {
printf("Usage: rsem-extract-reference-transcripts refName quiet gtfF sources hasMappingFile [mappingFile] chromosome_file_1 [chromosome_file_2 ...]\n");
exit(-1);
}
verbose = !atoi(argv[2]);
if (hasMappingFile) {
loadMappingInfo(argv[6]);
}
sources.clear();
if (strcmp(argv[4], "None")) parseSources(argv[4]);
parse_gtf_file(argv[3]);
ifstream fin;
string line, gseq, seqname;
int len;
size_t seqlen;
chrvec.clear();
seqs.clear();
seqs.resize(M + 1, "");
int start = hasMappingFile ? 7 : 6;
for (int i = start; i < argc; i++) {
fin.open(argv[i]);
general_assert(fin.is_open(), "Cannot open " + cstrtos(argv[i]) + "! It may not exist.");
cursor.filename = argv[i]; cursor.line_no = cursor.pos = 0;
getline(fin, line);
while ((fin) && (line[0] == '>')) {
istringstream strin(line.substr(1));
strin>>seqname;
++cursor.line_no;
gseq = ""; seqlen = 0;
while((getline(fin, line)) && (line[0] != '>')) {
++cursor.line_no;
len = line.length();
for (cursor.pos = 0; cursor.pos < len; ++cursor.pos) line[cursor.pos] = check(line[cursor.pos]);
seqlen += len;
gseq += line;
}
assert(seqlen > 0);
iter = sn2tr.find(seqname);
if (iter == sn2tr.end()) continue;
chrvec.push_back(ChrInfo(seqname, seqlen));
vector<int>& vec = iter->second;
int s = vec.size();
for (int j = 0; j < s; j++) {
assert(vec[j] > 0 && vec[j] <= M);
transcripts.getTranscriptAt(vec[j]).extractSeq(gseq, seqs[vec[j]]);
}
}
fin.close();
if (verbose) { printf("%s is processed!\n", argv[i]); }
}
sort(chrvec.begin(), chrvec.end());
shrink();
if (verbose) { printf("Extracting sequences is done!\n"); }
writeResults(argv[1]);
return 0;
}
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