A kind of Siniperca tlr7 gene and its application
A technology of mandarin mandarin fish and genes, which is applied in aquaculture and biological fields, can solve problems such as the report on gene expression research of TLR7 cloning pathway of mandarin mandarin fish, and achieve the effect of improving economic benefits
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Embodiment 1
[0025] 1. Cloning of TLR7 cDNA of Siniperca sinensis
[0026] The primers used for TLR7 gene cDNA amplification are shown in Table 1 below:
[0027] Table 1 The sequences of primers used in TLR7 gene cDNA amplification
[0028] Primer name Primer sequence (5'-3') Purpose GSP-TLR7-1 CAGTCCACCTCTCACCA TLR7 5'-Race GSP-TLR7-2 CTGCCGTTGCTGGTCACA TLR7 5'-Race GSP-TLR7-3 ATGGCAGGGTTTTTGGGT TLR7 5'-Race 3' CDS Primer A AAGCAGTGGTATCAACGCAGACTAC TLR7 3'-Race GSP-TLR7-4 GAAAGTGCCTTGCAATTCAAAGTACCTA TLR7 3'-Race GSP-TLR7-5 GTATAAGCGGTCTGTGCTGGAGTGG TLR7 3'-Race AUAP GGCCACGCGTCGACTAGTAC Universal 5'-Race AAP GGCCACGCGTCGACTAGTAC(G) 16
Universal 5'-Race UPM CTAATACGACTCACTATAGGGC Universal 3’-Race
[0029] 1. Amplification of Siniperca sinensis TLR7 5' end sequence
[0030] (1) Synthesis, purification and tailing of the first strand of cDNA
[0031] Use SUPERSCRIPT II RT enzyme and pr...
Embodiment 2
[0087] Infectious spleen and kidney necrosis virus (ISKNV) and Aeromonas hydrophila (Aeromonas hydrophila) are the most serious virus and bacterial pathogens in mandarin fish culture. Renal necrosis virus and Aeromonas hydrophila) were tested for real-time detection of TLR7-Myd88-dependent disease resistance signaling pathway-related genes (TLR7, Myd88, IRAK1 and The change of IRF7) expression is an auxiliary detection method for Siniperca sinensis infected by virus or bacteria.
[0088] Experimental method: A total of 100 healthy mandarin ducks with an average weight of 42.63g were placed in an aquarium with a water flow system for two weeks before the experiment began. The breeding conditions were continuous aeration, water temperature 26°C, Feed three times. Before the infection experiment, 10 fish were randomly selected to check the common pathogens such as infectious spleen and kidney necrosis virus (ISKNV) and Aeromonas hydrophila in the body, and after ensuring that th...
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