Highly effective polypeptide for inhibiting angiogenesis, physical chemistry modifying method and application thereof
A technology of angiogenesis and angiogenesis inhibition, which is applied in the direction of cardiovascular system diseases, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc., can solve the problems of insufficient vascular specificity and selectivity, production scale and Increased production costs, increased drug side effects, etc.
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[0039] 1. Cloning of RGD-ED gene and construction of its prokaryotic expression vector
[0040] The base encoding the RGD-ED polypeptide sequence was synthesized as a template; an upstream primer and a downstream primer were synthesized, wherein the upstream primer added an NdeI restriction site; the downstream primer contained an Arg-Gly-Asp sequence and an XhoI site. Perform PCR amplification, the amplified product is recovered and purified by agarose gel electrophoresis, digested with NdeI and XhoI, cloned into the prokaryotic expression vector pw, positive clones are screened by PCR, and the nucleotide sequence analysis confirms that the designed mutation has occurred in the sequence .
[0041] Synthetic primer 1: 5'GGAATTCCATATG ATCGTGCGCCGTGCCGACCGC3'
[0042] Synthetic primer 2: 5'CCGCTCGAGGCAGAAGCAGTCACCACGGCA3'
[0043] Among them, primer 1 encodes the partial sequence of NdeI site and Ile-Val-Arg-Arg-Ala-Asp-Arg-Ala-Ala-Val-Pro, primer 2 encodes XhoI site and conta...
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