Expressed sequence tag analysis for identification and characterization of sex-related genes in the giant tiger shrimp Penaeus monodon.
Sex-related genes expressed in vitellogenic ovaries of the giant tiger shrimp, Penaeus monodon, were identified by an EST approach. A total of 1051 clones were unidirectionally sequenced from the 5 terminus. Nucleotide sequences of 743 EST (70.7%) significantly matched known genes previously deposited in the GenBank (E-value <10(-4)) whereas 308 ESTs (29.3%) were regarded as newly unidentified transcripts (E-value >10(-4)). A total of 559 transcripts (87 contigs and 472 singletons) were obtained. Thrombospondin (TSP) and peritrophin (79 and 87 clones accounting for 7.5 and 8.3% of clones sequenced, respectively) predominated among characterized transcripts. Several full length transcripts (e.g. cyclophilin, profillin and thioredoxin peroxidase) were also isolated. A gene homologue encoding chromobox protein (PMCBX, ORF of 567 nucleotides encoding a protein of 188 amino acids) which is recognized as a new member of the HP1 family was identified. Expression patterns of 14 of 25 sex-related gene homologues in ovaries and testes of P. monodon broodstock were examined by RT-PCR. Female sterile and ovarian lipoprotein receptor homologues were only expressed in ovaries whereas the remaining transcripts except disulfide isomerase related P5 precursor and adenine nucleotide translocator 2 were higher expressed in ovaries than testes of P. monodon broodstock. A homologue of ubiquitin specific proteinase 9, X chromosome (Usp9X) revealed a preferential expression level in ovaries than testes of broodstock-sized P. monodon (N = 13 and 11, P <0.05) but was only expressed in ovaries of 4-month-old shrimp (N = 5 for each sex).
Preechaphol
Rachanimuk
R
Program in Biotechnology, Chulalongkorn University, Bangkok 10330, Thailand.
Leelatanawit
Rungnapa
R
Sittikankeaw
Kanchana
K
Klinbunga
Sirawut
S
Khamnamtong
Bavornlak
B
Puanglarp
Narongsak
N
Menasveta
Piamsak
P
D000595:Amino Acid Sequence; D000818:Animals; D002999:Clone Cells; D020224:Expressed Sequence Tags; D005260:Female; D015723:Gene Library; D005796:Genes; D008297:Male; D008969:Molecular Sequence Data; D010053:Ovary / Q000378:metabolism; D033561:Penaeidae / Q000235:genetics*; D010802:Phylogeny; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012727:Sex Characteristics; D012733:Sex Differentiation / Q000235:genetics*; D013737:Testis / Q000378:metabolism
D011506:Proteins; D012333:RNA, Messenger