Method for realizing fermentative expression of angiotensin converting enzyme 2 by using prokaryotic cell
A technology of prokaryotic organisms and extracellular regions, which is applied in the biological field to achieve the effects of large protein extraction, increased water solubility, and low price
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Embodiment 1
[0068] Embodiment 1 uses the design and result analysis of expressing ACE2 in Escherichia coli
[0069] The first step is to construct a recombinant expression vector with a HIS tag or a Strep-II tag.
[0070] Wherein, the ACE2 (18-805aa) target protein sequence is the sequence of SEQ ID NO.1;
[0071] The nucleotide sequence corresponding to the amino acid sequence of SEQ ID NO.1 is the sequence of SEQ ID NO.2;
[0072] ACE2 (18-740aa) target protein sequence is the sequence of SEQ ID NO.3;
[0073] The nucleotide sequence corresponding to the amino acid sequence of SEQ ID NO.3 is the sequence of SEQ ID NO.4;
[0074] ACE2 (18-615aa) target protein sequence is the sequence of SEQ ID NO.5;
[0075] The nucleotide sequence corresponding to the amino acid sequence of SEQ ID NO.5 is the sequence of SEQ ID NO.6;
[0076] The nucleotide sequence of the empty vector (derived from pET-30a(+)) used was set as SEQ ID NO.7.
[0077] It should be noted that the sequences inserted in...
Embodiment 2
[0129] Example 2 Production process and result analysis using Corynebacterium glutamicum to express ACE2
[0130] The first step is to construct a recombinant expression vector (with signal peptide)
[0131] In order to improve the expression effect, in particular a signal peptide sequence was also inserted before the ACE2 gene.
[0132] The plasmid constructed here is pXMJ19, and the involved signal peptide Sec signal peptide BZH001-cspB has the sequence SEQ ID NO.8:
[0133] ATGTTCAACAACCGTATCCGTACCGCCGCCCTCGCAGGTGCCATCGCAATCTCTACCGCAGCGTCTGGTCTGGTGGTCCCCAGCATTCGCTCAGGAA; alternatively, the TAT signal peptide CGR0949 is used, which has the sequence SEQ ID NO.9:
[0134] ATGCAAATAAACCGCCGAGGCTTCTTAAAAGCCACCGCAGGACTTGCCACTATCGGCGCTGCCAGCATGTTTATGCCAAAGGCCAACGCCCTTGGAGCA.
[0135] The second step is to select the expression host bacteria and introduce the vector
[0136] The selected strain was C. glutamicum CGMCC1.15647 (BZH001). Similar to Example 1, the engineering plasm...
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