Peptide for high performance inhibition of angiogenesis and method for preparing same and use thereof
An angiogenesis and high-efficiency technology, applied in biochemical equipment and methods, botanical equipment and methods, applications, etc., can solve the problems of increased difficulty in drug quality control, insufficient vascular specificity and selectivity, and ineffective targets. Clear and other problems, to achieve the effect of improving and inhibiting the growth of endothelial cells, improving and inhibiting the growth of endothelial cells, improving and anti-tumor effect
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[0034] 1. Cloning of RGD-ED gene and construction of its prokaryotic expression vector
[0035] 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 .
[0036] Synthetic primer 1: 5'GGAATTCCATATGATCGTGCGCCGTGCCGACCGC3'
[0037] Synthetic primer 2: 5'CCGCTCGAGGCAGAAGCAGTCACCACGGCA3'
[0038] 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 contai...
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