GAAP: Genome-organization-framework-Assisted Assembly Pipeline for prokaryotic genomes

BackgroundNext-generation sequencing (NGS) technologies have greatly promoted the genomic study of prokaryotes. However, highly fragmented assemblies due to short reads from NGS are still a limiting factor in gaining insights into the genome biology. Reference-assisted tools are promising in genome assembly, but tend to result in false assembly when the assigned reference has extensive rearrangements.ResultsHerein, we present GAAP, a genome assembly pipeline for scaffolding based on core-gene-defined Genome Organizational Framework (cGOF) described in our previous study. Instead of assigning references, we use the multiple-reference-derived cGOFs as indexes to assist in order and orientation of the scaffolds and build a skeleton structure, and then use read pairs to extend scaffolds, called local scaffolding, and distinguish between true and chimeric adjacencies in the scaffolds. In our performance tests using both empirical and simulated data of 15 genomes in six species with diverse genome size, complexity, and all three categories of cGOFs, GAAP outcompetes or achieves comparable results when compared to three other reference-assisted programs, AlignGraph, Ragout and MeDuSa.ConclusionsGAAP uses both cGOF and pair-end reads to create assemblies in genomic scale, and performs better than the currently available reference-assisted assembly tools as it recovers more assemblies and makes fewer false locations, especially for species with extensive rearranged genomes. Our method is a promising solution for reconstruction of genome sequence from short reads of NGS.

[1]  Jingfa Xiao,et al.  Comparative genomic analysis of Streptococcus suis reveals significant genomic diversity among different serotypes , 2011, BMC Genomics.

[2]  Jian Wang,et al.  SOAPdenovo2: an empirically improved memory-efficient short-read de novo assembler , 2012, GigaScience.

[3]  Pietro Liò,et al.  Short and long-term genome stability analysis of prokaryotic genomes , 2013, BMC Genomics.

[4]  Anat Kreimer,et al.  The evolution of modularity in bacterial metabolic networks , 2008, Proceedings of the National Academy of Sciences.

[5]  Minping Qian,et al.  Flexibility and Symmetry of Prokaryotic Genome Rearrangement Reveal Lineage-Associated Core-Gene-Defined Genome Organizational Frameworks , 2014, mBio.

[6]  Eduardo P C Rocha,et al.  Gene essentiality determines chromosome organisation in bacteria. , 2003, Nucleic acids research.

[7]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[8]  Timothy P. L. Smith,et al.  Reducing assembly complexity of microbial genomes with single-molecule sequencing , 2013, Genome Biology.

[9]  Pietro Liò,et al.  MeDuSa: a multi-draft based scaffolder , 2015, Bioinform..

[10]  Steven J. M. Jones,et al.  Abyss: a Parallel Assembler for Short Read Sequence Data Material Supplemental Open Access , 2022 .

[11]  Tao Jiang,et al.  AlignGraph: algorithm for secondary de novo genome assembly guided by closely related references , 2014, Bioinform..

[12]  Jun Yu,et al.  PGAP: pan-genomes analysis pipeline , 2011, Bioinform..

[13]  Jiayan Wu,et al.  Complete Genome Sequence of Streptococcus suis Serotype 14 Strain JS14 , 2011, Journal of bacteriology.

[14]  Brian J. Raney,et al.  Ragout—a reference-assisted assembly tool for bacterial genomes , 2014, Bioinform..

[15]  Jingfa Xiao,et al.  Complete Genome Sequence of Streptococcus suis Serotype 3 Strain ST3 , 2011, Journal of bacteriology.

[16]  D. Leach,et al.  Bacterial Genome Instability , 2014, Microbiology and Molecular Reviews.

[17]  E. Birney,et al.  Velvet: algorithms for de novo short read assembly using de Bruijn graphs. , 2008, Genome research.

[18]  Qing-Yong Yang,et al.  Spatial features for Escherichia coli genome organization , 2015, BMC Genomics.

[19]  E. Cox,et al.  Gene location and DNA density determine transcription factor distributions in Escherichia coli , 2012, Molecular systems biology.

[20]  William Stafford Noble,et al.  Three-dimensional modeling of the P. falciparum genome during the erythrocytic cycle reveals a strong connection between genome architecture and gene expression , 2014, Genome research.

[21]  Loretta Auvil,et al.  Reference-assisted chromosome assembly , 2013, Proceedings of the National Academy of Sciences.