Abstract
The cockroach Periplaneta americana is a notorious pest and threat to health worldwide, with a high reproductive ability. However, a limited amount of data is available on the developmental stage-specific transcriptomes of P. americana. To identify genes involved in developmental processes and to develop additional SSR markers in P. americana, we carried out de novo assembly of the P. americana transcriptome using Illumina sequencing. After removing low-quality sequences, we obtained 64,954,709 contigs, which were further assembled into 125,390 unigenes with an average length of 711 bp. Based on similarity searches against known proteins, we identified 48,300 unigenes based on a cut-off E-value of 10−5. The assembled sequences were annotated according to gene descriptions, gene ontology and clusters of orthologous groups. A total of 14,195 potential SSRs were identified, and 41 of 63 randomly chosen primer pairs successfully amplified the predicted SSR markers, seven of which were polymorphic in size in P. americana. Furthermore, the Spag6 gene was confirmed to be testes specific, and the fru and RPSA genes were related to the development of the testis. This is the special report of a P. americana transcriptome obtained using Illumina sequencing technology, and a large number of molecular markers were developed.
Highlights
The cockroach is one of the oldest known winged insects, and its habitats are closely associated with those of humans
We focus on two types of markers, simple sequence repeats (SSRs) and single-nucleotide polymorphisms (SNPs)
The Sperm-associated Antigen 6 (Spag6) gene was confirmed to be testes specific, and the fruitless and RPSA genes were related to the development of the testis
Summary
The cockroach is one of the oldest known winged insects, and its habitats are closely associated with those of humans. Next-generation sequencing has significantly improved the sensitivity of gene expression profiling and contributed to comparative genomic studies[8,9]. For non-model organisms with limited genomic information, transcriptome sequencing is a cost-effective tool because it focuses mostly on the sequencing of functional and protein-coding RNAs12. Transcriptome sequencing is an effective method for obtaining EST sequences, which are essential for developing molecular markers and identifying novel genes. Deep transcriptome sequencing provides a good resource for the development of SSRs because of its high throughput. Another type of marker, SNPs, are the most abundant type of marker and can be detected via high-throughput sequencing, which will be helpful in future linkage and associated studies
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