Nile tilapia NPB29 (neuropeptide B29) peptide as well as encoding nucleic acid and application thereof
A technology of Nile tilapia and tilapia, which is applied in the field of functional application of 29 peptide, can solve the problems that the functional research is still blank
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[0025] Below in conjunction with experimental data, further illustrate the present invention.
[0026] Cloning of NPB cDNA from Nile Tilapia
[0027] (1) Primer design
[0028] Using the EST retrieval function of NCBI, we found a part of the predicted sequence XM_003450122.1 published in tilapia, and then designed specific primers to clone partial fragments of NPB cDNA, and then designed 5`-RACE and Specific primers for 3'RACE, so as to carry out 5'-RACE and 3'RACE with the designed degenerate primers. . Finally, specific primers were designed in the 3' and 5' non-coding regions to clone the open reading frame ORF.
[0029] (2) Cloning of the middle fragment of Nile tilapia NPB cDNA
[0030] The first strand cDNA was synthesized from the total RNA using the Rever Tra Ace-a-Tm First Strand cDNA Synthesis Kit from Toyobo. The PCR amplification of the middle fragment was carried out according to the Toyobo product polymerase blend tap plus. The first round of PCR reaction c...
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