Study On MTOR Signaling Pathway In Regulating Rapid Fish Growth

The mTOR signaling pathway(AKT/mTOR/4E-BP1/S6K)plays an important role in cell growth,development,and protein synthesis.In this paper,diploid red crucian carp and triploid Xiangyun crucian carp were used as materials to conduct experiments on gene expression related to mTOR signaling pathway.In order to further study the relationship between the mTOR signaling pathway and the rapid growth of fish,we used yellow catfish with obvious males of the same age to grow faster than females,using gene cloning,RT-PCR,qRT-PCR and Western blotting techniques The main genes of mTOR signaling pathway AKT1/AKT2/AKT3/mTOR/S6KA/S6KB/4E-BP1 were cloned and expressed.The result is as follows:1.Experiments show that the expression of mTOR signaling pathway-related genes in 7 different tissues of liver,kidney,intestine,heart,gonad,brain and muscle of diploids red crucian carp and triploids has significant differences,except for 4E The expression of BP1 gene and mTOR / AKT1 / S6 KA gene in gonadal tissues and the expression of AKT1 gene in kidney tissue were significantly higher in diploid red crucian carp than in triploid Xiangyun crucian carp(P<0.05).Triploid Xiangyun crucian carp is significantly higher than diploid red crucian carp(P<0.05).2.The full length sequence of AKT2 / 4E-BP1 gene was obtained bygene cloning method,which were 1431 bp and 824 bp,respectively,and predicted to encode 476 and 262 amino acids.At the same time,a partial sequence of the AKT3/S6KA/mTOR/AKT1/S6 KB gene was also cloned,and blast alignment and phylogenetic tree analysis were performed.It was found that the above genes have high homology with other species,of which AKT2/4E-BP1 and homologous species The similarities are99.92% and 99.82%,respectively.3.The differential expression of AKT3 / S6 KB / AKT3 / S6 KA /S6 KB / 4E-BP1 / mTOR gene in 7 different tissues of male and female Pelteobagrus fulvidraco was detected by qRT-PCR.It was found that the expression of 4E-BP1 gene in gonad tissue is that the expression level in males is lower than that in female(P<0.05),and the expression of S6 KA gene in heart and brain tissue is lower than that of females in males(P<0.05).Except for gonad tissue,the expression of the other 6genes in male Pelteobagrus fulvidraco was significantly higher than that of female Pelteobagrus fulvidraco(P<0.05).4.The results of western blotting showed that mTOR gene protein was expressed in the muscle and intestine tissues of Pelteobagrus fulvidraco,while the expression in the kidney tissue was weak.