Author(s)

Ruocheng Du ¹

Affiliation(s)

¹ RCF Experimental School, Beijing, China

Corresponding Author

Ruocheng Du

ABSTRACT

In this study, we employed molecular cloning techniques involving PCR, DNA agarose gel electrophoresis, and restriction enzyme digestion to successfully clone 2201LBD mutants with point mutations at residues 172 and 142. Following enzymatic digestion of DNA and plasmid vectors using restriction enzymes, the T4 DNA ligase was used to ligate the modified DNA fragments into the vectors. Subsequently, the resulting constructs were transformed into Escherichia coli BL21 (DE3) strains for protein expression and purification. These engineered mutants, 2201LBD-Y172A and 2201LBD-R142A, were purified to facilitate subsequent experimental investigations.

KEYWORDS

Gene cloning construction, PCR, T4 DNA ligase, agarose gel electrophoresis stands, protein purification

CITE THIS PAPER

Ruocheng Du, Cloning and Protein Purification of 2201LBD Mutants. MEDS Basic Medicine (2023) Vol. 1: 26-31. DOI: http://dx.doi.org/10.23977/medbm.2023.010205.

REFERENCES

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