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Turkish Journal of Fisheries and Aquatic Sciences 2014, Vol 14, Num, 3     (Pages: 689-696)

Molecular Characterization, Genomic Organization, and Expression Analysis of Sperm Gelatinase Gene during Post-Embryonic Development in Macrobrachium nipponense

Shiyong Zhang 1 ,Hongtuo Fu 1-2 ,Wenyi Zhang 2 ,Hui Qiao 2 ,Shengming Sun 2 ,Hongkun Bai 1 ,Yiwei Xiong 2 ,Sufei Jiang 2 ,Yongsheng Gong 2 ,Shubo Jin 1-2

1 Wuxi Fishery College Nanjing Agricultural University, 214081, Wuxi, China
2 Chinese Academy of Fishery Sciences, Ministry of Agriculture, Freshwater Fisheries Research Center, Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, 214081, Wuxi, China
DOI : 10.4194/1303-2712-v14_3_11 Viewed : 4203 - Downloaded : 2824 Crustacean sperm gelatinase first identified in Macrobrachium rosenbergii encodes a gelatinolytic activity protease belonging to a kind of sperm proteases and probably involve in the fertilization process, which could be inhibited by a male reproductive tract-specific peptidase inhibitor (MRPINK) specifically. In this study, we have isolated the full-length cDNA and its genomic DNA sequence of a sperm gelatinase homologue from the testis of oriental river prawn (Macmbrachium nipponense) according to the established expressed sequence tags (ESTs) information using Rapid Amplification of the cDNA Ends (RACE) technique, designated as MnSG. The full-length cDNA of MnSG is 575bp in size and has an open reading frame (ORF) of 384bp, encoding a 127-amino acid protein. Genomic DNA sequence contains 1 intron and 2 exons, and three microsatellite sequences, (ATAC)5(AC)13(AT)15, (TA)11, AT)4, (AT)11, (AT)11 and (TA)37, were identified in the intron. Real-time quantitative PCR analyses revealed that MnSG expresses specifically in the testis, while genomic DNA of MnSG exists in both male and female prawn. The expression level increase gradually during growth and development process of male juvenile prawn and sharply from PL30 to PL40. Keywords : Macrobrachium nipponense, sperm gelatinase, sperm protease, fertilization, autumn propagation