title: Identification of sex-specific expression markers in the giant tiger shrimp (Penaeus monodon).
abstract: Bulked segregant analysis (BSA) and AFLP were used for isolation of genomic sex determination markers in Penaeus monodon. A total of 256 primer combinations were tested against 6-10 bulked genomic DNA of P. monodon. Five and one candidate female- and male-specific AFLP fragments were identified. Female-specific fragments were cloned and further characterized. SCAR markers derived from FE10M9520, FE10M10725.1, FE10M10725.2 and FE14M16340 provided the positive amplification product in both male and female P. monodon. Further analysis of these markers using SSCP and genome walk analysis indicated that they were not sex-linked. In addition, sex-specific (or differential) expression markers in ovaries and testes of P. monodon were analyzed by RAP-PCR (150 primer combinations). Twenty-one and fourteen RAP-PCR fragments specifically/differentially expressed in ovaries and testes of P. monodon were successfully cloned and sequenced. Expression patterns of 25 transcripts were tested against the first stranded cDNA of ovaries and testes of 3-month-old and broodstock-sized P. monodon (N=5 and N=7-10 for females and N=4 and N=5-7 for males, respectively). Five (FI-4, FI-44, FIII-4, FIII-39 and FIII-58) and two (M457-A01 and MII-51) derived RAP-PCR markers revealed female- and male-specific expression patterns in P. monodon. Surprisingly, MII-5 originally found in testes showed a higher expression level in ovaries than did testes of juvenile shrimps but a temporal female-specific pattern in P. monodon adults.
authors.affiliation: Center of Excellence for Marine Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Bavornlak
authors.initials: B
authors.lastname: Khamnamtong
authors.forename: Supaporn
authors.initials: S
authors.lastname: Thumrungtanakit
authors.forename: Sirawut
authors.initials: S
authors.lastname: Klinbunga
authors.forename: Takashi
authors.initials: T
authors.lastname: Aoki
authors.forename: Ikuo
authors.initials: I
authors.lastname: Hirono
authors.forename: Piamsak
authors.initials: P
authors.lastname: Menasveta
mesh_terms: D000818:Animals; D001483:Base Sequence; D005260:Female; D015870:Gene Expression; D005819:Genetic Markers; D008297:Male; D010053:Ovary / Q000502:physiology; D033561:Penaeidae / Q000235:genetics* / Q000502:physiology*; D012150:Polymorphism, Restriction Fragment Length; D013737:Testis / Q000502:physiology
chemical_list: D005819:Genetic Markers