Suppression subtractive hybridization (SSH) for isolation and characterization of genes related to testicular development in the giant tiger shrimp Penaeus monodon

被引:30
|
作者
Leelatanawit, Rungnapa [2 ]
Klinbunga, Sirawut [1 ,4 ]
Aoki, Takashi [5 ]
Hirono, Ikuo [5 ]
Valyasevi, Rudd [4 ]
Menasveta, Piamsak [1 ,3 ]
机构
[1] Chulalongkorn Univ, Fac Sci, Ctr Excellence Marine Biotechnol, Bangkok 10330, Thailand
[2] Chulalongkorn Univ, Fac Sci, Program Biotechnol, Bangkok 10330, Thailand
[3] Chulalongkorn Univ, Fac Sci, Dept Marine Sci, Bangkok 10330, Thailand
[4] Natl Sci & Technol Dev Agcy, Natl Ctr Genet Engn & Biotechnol BIOTEC, Pathum Thani 12120, Thailand
[5] Tokyo Univ Marine Sci & Technol, Grad Sch Marine Sci & Technol, Lab Genome Sci, Tokyo 1088477, Japan
关键词
Dmc1; Inx2; PGMRC1; Real-time PCR; SSH;
D O I
10.5483/BMBRep.2008.41.11.796
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 A 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 A 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 A 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). [BMB reports 2008; 41(11): 796-802]
引用
收藏
页码:796 / 802
页数:7
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