Suppression subtractive hybridization (SSH) for isolation and characterization of genes related to testicular development in the giant tiger shrimp Penaeus monodon.
Suppression subtractive hybridization (SSH) cDNA libraries of the giant tiger shrimp, Penaeus monodon, were constructed. In total, 178 and 187 clones from the forward and reverse SSH libraries, respectively, of P. monodon were unidirectionally sequenced. From these, 37.1% and 53.5% Expressed Sequence Tags (ESTs) significantly matched known genes (E-value < 1e-04). Three isoforms of P. monodon progestin membrane receptor component 1: PM-PGMRC1-s (1980 bp), PM-PGMRC1-m (2848 bp), and PM-PGMRC1-l (2971 bp), with an identical ORF of 573 bp corresponding to a deduced polypeptide of 190 amino acids, were successfully identified by RACE-PCR. Interestingly, PMPGMRC1 showed a greater expression level in testes of juvenile than broodstock P. monodon (P < 0.05). Dopamine administration (10(-6) mol/shrimp) resulted in up-regulation of PMPGMRC1 in testes of juveniles at 3 hrs post treatment (P < 0.05), but had no effect on PM-Dmc1 (P > 0.05).
Leelatanawit
Rungnapa
R
Program in Biotechnology, Chulalongkorn University, Bangkok, Thailand.
Klinbunga
Sirawut
S
Aoki
Takashi
T
Hirono
Ikuo
I
Valyasevi
Rudd
R
Menasveta
Piamsak
P
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D055028:Comparative Genomic Hybridization / Q000379:methods*; D020224:Expressed Sequence Tags; D015723:Gene Library; D005796:Genes; D008297:Male; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D013737:Testis / Q000254:growth & development* / Q000378:metabolism