The androgenic gland (AG), playing an important role in sex differentiation of male crustacean, is a target candidate to understand the mechanism of male development and to mine male-specific sex markers. An SSH library (designated as male reproduction-related tissues—SSH library, MRT-SSH library for short) was constructed using cDNA from tissues located at the basal part of the 5th pereiopods, including AG and part of spermatophore sac, as tester, and the cDNA from the basal part of the 4th pereiopods of these male shrimp as driver. 402 ESTs from the SSH library were sequenced and assembled into 48 contigs and 104 singlets. Twelve contigs and 14 singlets were identified as known genes. The proteins encoded by the identified genes were categorized, according to their proposed functions, into neuropeptide hormone and hormone transporter, RNA posttranscriptional regulation, translation, cell growth and death, metabolism, genetic information processing, signal transduction/transport, or immunity-related proteins. Eleven highly expressed contigs in the SSH library were selected for validation of the MRT-SSH library and screening sex markers of shrimp. One contig, specifically expressed in male shrimp, had a potential to be developed as a transcriptomic sex marker in shrimp. 1. Introduction The existence of sex dimorphism is a very common phenomenon in crustaceans. A number of crustacean species exhibit bimodal growth patterns in which males exhibit superior growth to females or vice versa [1]. Female penaeid shrimps usually show superior growth than males when they reach maturation. For example, females exhibit approximately 10–20% higher growth rate than males in Penaeus monodon [2]. Therefore, culture of monosex population is a good way to raise the production of cultured species. Understanding the mechanism of sex determination and sex differentiation will facilitate the development of molecular tools for producing monosex populations in crustacean species [3]. However, little information about sex determination cascades and sexdiagnostic markers is available in penaeid shrimp [4]. A ZW sex determination system in penaeid shrimp was proposed based on the data that the progeny of interspecific hybridization between females of P. monodon and males of Penaeus esculentus had a higher proportion (86%) of males than females [5]. The following reports that female specific-markers were found in the female genetic linkage maps for Penaeus japonicus [6] and Litopenaeus vannamei [7] and that the induced triploids P. japonicus were always female [8, 9] also support
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