F MnFtz-f1 have been Lipoxygenase web compared with those of other crustaceans by DNAMAN
F MnFtz-f1 were compared with those of other crustaceans by DNAMAN six.0. The results showed that MnFtz-f1 had significant homology with Ftz-f1 of other crustaceans, and each had the DNA-binding domain (DBD) and activation factor-2 (AF-2) as conserved domains. MnFtz-f1 showed the highest amino acid identity (68.3 ) with Ftz-f1 of Penaeus vannamei followed by Penaeus monodon (68.1 ) and Homarus americanus (50.2 ) (Figure two). A phylogenetic tree of insects and crustaceans was constructed by MEGA five.1 computer software. The results showed that the amino acid sequence of H. americanus clustered together with the amino acid sequence of MnFtz-f1. The phylogenetic tree was clearly divided into two main branches, i.e., insects and crustaceans (Figure three). The iterative threading assembly refinement (I-TASSER) server (42, 43) was made use of to analyze and evaluate the Ftz-f1 amino acid sequences of M. nipponense and also other crustaceans. The outcomes with the three-dimensional (3D) atom model generated by I-TASSER showed that the Ftz-f1 amino acid sequences of M. nipponense, P. vannamei, and other crustaceans possess the similar DNA-binding domain (Figure four).Effect of 20E on the Expression of MnFtz-fThe expression degree of MnFtz-f1 inside the ovary below diverse concentrations of 20E was detected by qPCR (Figure 8). In comparison with the handle group, a low concentration of 20E (3 mg/g) had no substantial effect around the expression of MnFtz-f1 (P 0.05). When the concentration of 20E was five mg/g, the expression of MnFtz-f1 decreased drastically (P 0.05). The expression of MnFtz-f1 was considerably inhibited below the action of a higher concentration of 20E (20 mg/g) (P 0.05). The expression degree of MnFtz-f1 at unique time points was detected in the identical 20E concentration of 5 mg/g. The results showed that compared to the control group, the expression level of MnFtz-f1 was significantly decreased just after 20E administration (P 0.05). MnFtz-f1 expression decreased to the lowest level at 12 h after which enhanced progressively.Effect of MnFtz-f1 Gene Knockdown on the Expression of MnFtz-f1, Vg, Mn-Spook, and Phantom in the OvaryThe function of MnFtz-f1 in M. nipponense and its regulatory relationship with other genes had been studied by the RNAi process (Figure 9). In comparison with the manage group, the expression level of MnFtz-f1 did not reduce significantly within 24 h right after dsMnFtz-f1 RNA administration (P 0.05). The expression degree of MnFtz-f1 at 48 and 96 h after the administration was considerably decreased by 97.12 and 86.09 , respectively, as compared to that with the control group (P 0.05). Immediately after silencing of MnFtz-f1, the expression levels of Mn-Spook, Phantom, and Vg decreased substantially at 48 and 96 h following the administration, plus the levels decreased by 51.42 and 66.06 , 56.16 and 69.82 , and 77.14 and 79.50 , SSTR2 Storage & Stability respectively (P 0.05).Expression from the MnFtz-f1M Gene in Distinctive TissuesThe distribution of MnFtz-f1 gene expression in different tissues (ovary, heart, gill, eyestalk, hepatopancreas, and muscle) of M. nipponense was determined by qPCR. As shown in Figure five, the highest mRNA expression was observed in the ovary, followed by that in the heart (P 0.05). The expression levels of MnFtz-f1 in the ovary, heart and gill had been 57.5-fold, 11.8-fold, and 6.2-fold larger than that in the muscle, respectively.Expression with the MnFtz-f1 Gene in Diverse Developmental Stages on the OvariesAs shown in Figure six, the expression level of MnFtz-f1 mRNA was the highest within the O2 stage and t.