| Tilapia is one of the world’s major aquaculture object. It is one excellent farmed fish which the FAO introduced to the whole world. The production of farmed tilapia in China ranks first in the world and Nile tilapia is the most widely breeding species. There is a big difference between the male and female Nile tilapia in size, growth rate, etc. The growth rate of males is about40-50%faster than females and the sexually mature males was significantly greater than the individual females, which seriously affecting the overall commercial fish out of the pool, reducing the yield and quality of Nile tilapia. In addition, as breed quickly, mature early, Nile tilapia in the breeding process prone to breeding density is too large, growth is blocked, malnutrition, fish is too small and some other unfavorable circumstances, which directly affect the market size tilapia. Gender-specific molecular markers combines with the all-male tilapia cultivation technology is better to improve the efficiency of all-male tilapia breeding.In the experiment screening of the gender differentially expressed genes between male and female Nile tilapia muscle tissue, Oreochromis niloticus, eight differentially expressed genes were identified and sequenced by amplification with20random primers. Sequences analysis showed that five of eight differentially expressed genes were60S ribosomal protein L3(RL3), muscle-type creatine kinase M2-CK, Parvalbumin beta-likeand transcription factor Sox4, transcription variant3(LOC100691543) genes, respectively. The rest three genes was unknown. RT-PCR analysis showed that the expression of8differentially expressed genes in Nile tilapia male and female muscle tissue were significant different. Base on quantitative Real-time PCR, the expression level of ACP3-X, ACP15-X,60S ribosomal protein L3(RL3), muscle-type creatine kinase M2-CK, Parvalbumin beta-likeand transcription factor Sox4gene in female Nile tilapia muscle were significantly higher than in male fish(P<0.01), while the expressions of ACP6-Y and transcription variant3(LOC100691543) genes in male Nile tilapia were significantly higher than in female fish(P<0.01). All the8ESTs were screened by annealing control primer system may participate in regulating on muscle growth of male and female Nile tilapia. This study established a foundation for further screening of muscle growth related genes.Nile tilapia females (XX) genome difference between gene screening experiment, select the XX, YY genotype Nile tilapia for each10caudal mixed pools of genomic DNA samples formed to female fish genomic Tester DNA, ultra-male genomic DNA as the driver, using suppression subtractive hybridization technique to build Nile tilapia females subtractive hybridization library of the genomic DNA. Sequencing1000clone randomly selected from the library, obtained942ESTs. A total of125comments to the results of the sequence, a sequence of max1229bp, a minimum of157bp.125already known genes including calsequestrin1, insulin-like growth factor1receptor-like,60S ribosomal protein L36a, and other genes. GO-Fuction of all uniESTs in Nile tilapia females (XX) subtractive library show that molecular functional classification related ESTs most distributed in binding and catalytic activity, Cellular Component related ESTs most distributed in cell part and cell, Biological Process related ESTs most distributed in cellular process and metabolic process. KEGG pathway analysis show that the subtractive hybridization library of genes involved in the ubiquitin-proteasome pathway, the insulin signaling pathway, and other metabolic processes. This study is the foundation for further study of Nile tilapia females (XX) specific molecular markers.YY genotype Nile tilapia genome difference between gene screening experiment, select the XX, YY genotype Nile tilapia for each10caudal mixed pools of genomic DNA samples formed to ultra-males genomic Tester DNA, female fish genomic DNA as the driver, using suppression subtractive hybridization technique to build Nile tilapia ultra-males subtractive hybridization library of the genomic DNA. Sequencing1000clone randomly selected from the library, obtained924ESTs. A total of131comments to the results of the sequence, a sequence of max1270bp, a minimum of166bp.131already known genes including myosin-VIIa-like, growth differentiation factor6-a-like, utrophin, kinesin-like protein KIF15-A-like, and other genes. GO-Fuction of all uniESTs in YY genotype Nile tilapia subtractive library show that molecular functional classification related ESTs most distributed in binding and catalytic activity, Cellular Component related ESTs most distributed in cell part and cell, Biological Process related ESTs most distributed in cellular process and metabolic process. KEGG pathway analysis show that the genes of subtractive hybridization library involved in the ABC transporters, TGF-beta signaling pathway, Calcium signaling pathway, and other metabolic processes.This study is the foundation for further study of YY genotype Nile tilapia specific molecular markers. |
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