Agkistrodon halys ussuriensis disintegrin site-directed mutants cDNA clone, expression and protein preparation method and application thereof
A technology for removing integrins and white-browed pit vipers, applied in the biological field, can solve the problems of difficult protein realization and great influence on human body, and achieve the effects of high safety, low side effects and high application value.
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[0063] The cDNA sequence of the site-directed mutant of Agkistrodon halys disintegrin, mutating A145, T146, and G148 in the cDNA sequence of Agkistrodon halys disintegrin into C145, C146, and A148, the obtained cDNA contains 219 base pairs, and the nucleotide sequence for:
[0064] GAGGCCGGAG AAGAATGTGA CTGTGGCTCT CCTGGAAATC CGTGCTGTGA 50
[0065] TGCTGCAACC TGTAAACTGA GACAAGGAGC ACAGTGTGCA GAAGGACTGT 100
[0066] GTTGTGACCA GTGCAGATTT ATGAAAAAAG GAACAGTATG CCGGCCAACA 150
[0067] AGGGGTGATG ACATGGATGA TTACTGCAAT GGCATATCTG CTGGCTGTCC 200
[0068] CAGAAATCCCTTCCATGCC 219
[0069] The encoded protein expressed by the site-directed mutant cDNA of Agkistrodon halys disintegrin consists of 73 amino acids, and its Ile49 and Ala50 are replaced by Pro49 and Thr50. The amino acid sequence is as follows:
[0070] Glu Ala Gly Glu Glu Cys Asp Cys Gly Ser Pro Gly Asn Pro Cys Cys Asp
[0071] 1 5 10 15
[0072] Ala Ala Thr Cys Lys Leu Arg Gln Gly Ala Gln Cys Ala Glu Gly Leu Cys
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