C1q receptor ptgc1qr gene of Portunus trituberculatus and its encoded protein and application
A technology of swimming crab trituratus and gene encoding, which is applied in the field of molecular biology and can solve the problems such as the report of the C1q receptor gC1qR of the swimming crab trituratus that has not yet been found.
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Embodiment 1
[0031] The PtgC1qR gene of Portunus trituberculatus C1q receptor is the base sequence shown in SEQ ID No.1.
[0032] The sequence listing SEQ ID No.1 is:
[0033]tctcacttctcccttaattttcgcccttatttgtggtcccagcgccctaaaactccctggaatgttgcgtggagccctgagccgggggttggtgtcctcgacgtgtcttcaggggcgtgtcatgtccctaaatacgccaataattagaaatattgccctagtgtcctgttgttctcctccatccccgggccggccttcctcttcctcttcttcttgccctccacgcctcgccacgccctccaagctgtgttcctgtggctgtggcattcacggcatccacacgagaggtgaccgggagctggtggagttcctgcaggaggagatcgtggcggagaggaagactttgcagccaaaccttccctcacaccttggggactttgctgtcaaggggaccagtgccgagctcaccctctcccgctccttccatgatgaaaaaatcaccatcaccctcaacgtgaaccacaccgtcgaccaggaggatgaaggcagtgctgagctgacccaggaccagacggaggcactgctgaagagtcggccaagcttcgaggtggacatagtgatcggagccaagacactctcctttacctgctcctttgtcggcccagcggaggtgcagggcgggcaggaggaagatgtgtttggcatcaacgaactgaccatctatgaaggggagtggtccgaggcaacctattgtgtgtctggggatattctcgatgggatgatgtacgacctactaatgaacatgctggaggaacgaggggtcactaatgaatttgccgaacagctcagcactctctgttctgagtatgagcactctctctatgtgggcct...
Embodiment 2
[0065] The base sequence of Portunus trituberculatus C1q receptor PtgC1qR is described in SEQ ID No. 1 in the sequence listing, and the amino acid sequence is described in SEQ ID No. 2 in the sequence listing.
[0066] The sequence listing SEQ ID No.2 is:
[0067] MLRGALSRGLVSSTCLQGRVMSLNTPIIRNIALVSCCSPPSPGRPSSSSSSCPPRLATPSKLCSCGCGIHGIHTRGDRELVEFLQEEIVAERKTLQPNLPSHLGDFAVKGTSAELTLSRSFHDEKITITLNVNHTVDQEDEGSAELTQDQTEALLKSRPSFEVDIVIGAKTLSFTCSFVGPAEVQGGQEEDVFGINELTIYEGEWSEATYCVSGDILDGMMYDLLMNMLEERGVTNEFAEQLSTLCSEYEHSLYVGLLQRLQDFVKRK
[0068] It has a complete coding protein containing 268 amino acids, the coding sequence has no signal peptide, the predicted molecular weight is 29.36kDa, and the isoelectric point is 4.81. The mature peptide has a typical 33 kDa mitochondrial acidic matrix protein (MAM33p) domain.
[0069] The specific operation for obtaining the PtgC1qR recombinant protein of Portunus trituberculatus C1q receptor is as follows:
[0070] According to the cDNA seque...
Embodiment 3
[0072] 1. Phenoloxidase inhibition test of PtgC1qR recombinant protein of Portunus trituberculatus:
[0073] Preparation of Portunus trituberculatus blood cell crush supernatant (HLS): extract blood cells from Portunus trituberculatus, quickly add to anticoagulant, and centrifuge for 10 minutes to obtain blood cell pellet. After the pellet was resuspended with 0.1M PBS (pH=7.0), it was beaten 4 times with 20kHz / 100W ultrasonic waves, and after each 30s, centrifuged at 12,000rpm at 4 degrees for 10min to remove the pellet. The supernatant is the blood cell lysis supernatant (HLS), and put it in a 4°C refrigerator for later use.
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