Two kinds of novel angiogenesis-inhibiting medicine and its uses in preventing and treating tumour
An angiogenesis inhibitory, drug-based technology, applied in the field of molecular pharmacology
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[0028] (1) Construction of rAAV-kallistatin and rAAV-vasostatin
[0029] The cDNAs of kallistatin and vasostatin were obtained by PCR, and the correctness was verified by DNA sequencing. The sequence of Kallistatin is identical to that reported by NCBI except for two nucleotides. Its two different nucleotides are listed at positions 1145 and 1146, resulting in a mutation, the original threonine becomes serine ( figure 1 A). Because vasostatin is a fragment of calreticulin, after the 180th codon of calreticulin, a stop codon (TAG) is introduced through the 3' primer, and truncated calreticulin, namely vasostatin, is generated after translation. The sequence of vasostatin is identical to the nucleotides 54-594 of calreticulin reported by NCBI.
[0030] In order to obtain constitutive high expression of kallistatin and vasostatin, the two cDNAs were respectively inserted between the ITRs of the AAV vector, and the promoters were CMV enhancer / chicken β-actin promoter. To enhan...
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