ReproArrayGTS: A cDNA microarray for identification of reproduction-related genes in the giant tiger shrimp Penaeus monodon and characterization of a novel nuclear autoantigenic sperm protein (NASP) gene

Apr 21, 2010Comparative biochemistry and physiology. Part D, Genomics & proteomics

Gene array for finding reproduction-related genes in giant tiger shrimp and study of a new sperm protein gene

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Abstract

A cDNA microarray containing 4992 features was used to analyze gene expression in the ovaries and testes of giant tiger shrimp (Penaeus monodon).

Transcripts were differentially expressed during four stages of ovarian development in P. monodon.
The nuclear autoantigenic sperm protein (NASP) gene was characterized and found to be 2126 bp in length.
An open reading frame of 1812 bp corresponded to a deduced protein of 603 amino acids for PmNASP.
Higher transcript levels of PmNASP were confirmed at later stages of ovarian development through quantitative real-time PCR.
The microarray platform demonstrated effectiveness for high-throughput screening of genes related to ovarian development.

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Expressed sequence tags (ESTs) were established from various tissues of the giant tiger shrimp (Penaeus monodon). To simultaneously examine expression patterns of a large number of transcripts in ovaries and testes of P. monodon, a cDNA microarray (ReproArray(GTS)) containing 4992 features amplified from cDNAs of ovary (1920) and testis (3072) EST libraries was constructed and subjected to high-throughput gene expression analysis in four different stages of ovarian development (previtellogenesis, vitellogenesis, early cortical rod and late cortical rod stages). Several transcripts were found to be differentially expressed during P. monodon ovarian development. Among many important reproduction-related genes with differential expression from microarray data, nuclear autoantigenic sperm protein (NASP) was further characterized by RACE-PCR. The full-length cDNA of P. monodon NASP (PmNASP) was 2126 bp in length containing an open reading frame (ORF) of 1812 bp corresponding to a deduced protein of 603 amino acids with 5? and 3?UTRs of 93 and 202 bp (excluding the poly A tail), respectively. Higher PmNASP transcript levels at later stages of ovarian development was consistently confirmed by quantitative real-time PCR. This study indicated that ReproArray(GTS) is effective for high-throughput screening of genes that play important roles in ovarian development of P. monodon.

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ReproArrayGTS: A cDNA microarray for identification of reproduction-related genes in the giant tiger shrimp Penaeus monodon and characterization of a novel nuclear autoantigenic sperm protein (NASP) gene

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