High level stable expression of recombinant HIV gp120 in glutamine synthetase gene deficient HEK293T cells.
Protein Expr Purif
; 181: 105837, 2021 05.
Article
in English
| MEDLINE | ID: covidwho-1057206
ABSTRACT
Due to the important pathological roles of the HIV-1 gp120, the protein has been intensively used in the research of HIV. However, recombinant gp120 preparation has proven to be difficult because of extremely low expression levels. In order to facilitate gp120 expression, previous methods predominantly involved the replacement of native signal peptide with a heterologous one, resulting in very limited improvement. Currently, preparation of recombinant gp120 with native glycans relies solely on transient expression systems, which are not amendable for large scale production. In this work, we employed a different approach for gp120 expression. Besides replacing the native gp120 signal peptide with that of rat serum albumin and optimizing its codon usage, we generated a stable gp120-expressing cell line in a glutamine synthetase knockout HEK293T cell line that we established for the purpose of amplification of recombinant gene expressions. The combined usage of these techniques dramatically increased gp120 expression levels and yielded a functional product with human cell derived glycan. This method may be applicable to large scale preparation of other viral envelope proteins, such as that of the emerging SARS-CoV-2, or other glycoproteins which require the presence of authentic human glycans.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
HIV Envelope Protein gp120
/
HIV-1
/
Glutamate-Ammonia Ligase
Limits:
Animals
/
Humans
Language:
English
Journal:
Protein Expr Purif
Journal subject:
Molecular Biology
Year:
2021
Document Type:
Article
Affiliation country:
J.pep.2021.105837
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