Artificial gene encoding a chimeric protein of human angiogenin, chimeric plasmids pJZZ-A, providing for the expression of the chimeric protein gene human angiogenin in Escherichia coli and a strain of Escherichia coli BL21 (DE3) / pJZZ-A-PRODUCER recombinant chimeric protein human angiogenin

摘要

1. An artificial gene encoding a human angiogenin himernyybelok has the complete nucleotide sequence and the corresponding amino acid sequence shown in FIG. 2.2. The chimeric plasmid pJZZ-A, providing expression of the artificial gene fusion protein ZZ-hAng in Escherichia coli, having a size of 4756 base pairs (bp), a molecular weight of 3086 kDa and composed in accordance with the physical and genetic map of the plasmid shown in FIG. 1: a) the synthetic gene of recombinant ZZ-Ang (positions: 1509 - 2267 bp) encoding a chimeric polypeptide comprising from N-end amino acid sequence of two IgG-binding domains (ZZ) of protein a Staphylococcus aureus, and the C- the end of the amino acid sequence of mature human angiogenin, and VC Luciano following genetic elements: - Lider and ZZ-region - 5'-terminal fragment (positions: 1509 - 1886 bp), a synthetic gene ZZ-Ang, encoding the amino acid sequence of the artificial leader of 8 amino acid residues and two IgG-binding domains (ZZ) of protein A; - hAngiogenin - 3'-terminal fragment (positions 1893 - 2264 bp), a synthetic gene ZZ-Ang, encoding the amino acid sequence of mature human angiogenin b) NdeI / XhoI fragment pJexpress414 plasmid comprising the elements: - pUC ori - a plasmid pUC replication site (positions 2 - 805 bp.) - terminato s transcription of various genes: terminator rpn txn (positions 977 - 1090 bp) terminator bla txn (positions: 1097 - 1397 bp), the terminator of bacteriophage T7 (position: 2284-2330 bp) terminator rrnB1 B2 T1 txn (positions: 2378 - 2552 bp.); - T7 promoter - T7 bacteriophage promoter (positions 1423 - 1439 bp.); - Ampicilin - β-lactamase gene that determines the stability of the bacteria to the beta lactam antibiotics (penicillin