A method for identifying Haitian melons using a specific peptide group
A technology of Haitian melon and sea cucumber, applied in the biological field, can solve the problems of lack of technical level and deceiving consumers.
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Embodiment 1
[0039] Example 1, Sequence Source and Alignment Information of Specific Polypeptides
[0040] 1. Haitian melon's unique source of peptides VMTDEMER, FNENEEMYEK, FNENEEMYETK, MSELESQVVSSK
[0041] The unique peptides VMTDEMER, FNENEEMYEK, FNENEEMYETK, and MSELESQVVSSK of Haitian melon all come from the protein tropomyosin [Apostichopus japonicus], and its Accesion number on NCBI is gi|302340969. The sequence is shown in SEQ ID NO.6 below. The first Glu was mutated into Val, the fourth Glu was mutated into Asp, and the polypeptide VMTDEMER was obtained; the second Thr of FTENEEMYEK was mutated into Asn, and the polypeptide FNENEEMYEK was mutated; the second Thr of FTENEEMYEKK was mutated into Asn, The 10th Lys was mutated into Thr, and the polypeptide FNENEEMYETK was obtained; the 6th Thr of MSELETQVMSSK was mutated into Ser, the 9th Met was mutated into Val, and the polypeptide MSELESQVVSSK was obtained; after NCBI blast comparison, the above polypeptide VMTDEMER , FNENEEMYEK,...
Embodiment 2
[0069] Embodiment 2, sea cucumber sample processing and detection steps
[0070] The steps of analyzing the sea cucumber samples to be tested include
[0071] (1) Sample pretreatment steps:
[0072] (1) Weigh 1g sea cucumber sample and homogenize it into a powder state, add 10mL protein extract (8M urea, 50mM NH 4 HCO 3 ) shake to extract protein, centrifuge at high speed and low temperature (20000r / min), take 200ul of supernatant and transfer to 1ml EP tube;
[0073] (2) Take 2μL of 1mol / L DTT (0.154g of dithiothreitol DTT dissolved in 1mL of deionized water) and add it to 100μL of the above protein solution and react at 37°C for 1 hour;
[0074] (3) Take 10 μL of freshly prepared IAA (accurately weigh 0.185g of iodoacetamide IAA and dissolve it in 1mL of deionized water to prepare a 1mol / L IAA mother solution for later use. The preparation process needs to be protected from light, and it is ready to use) and added to In the above reaction solution that has been cooled to...
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