A method for improving the thermal stability of interleukin il-34

Abstract

The invention relates to a method for improving the thermal stability of interleukin IL-34, and belongs to the field of biotechnology. The method comprises the steps: carrying out site-specific mutagenesis of a codon agc encoding No.98 serine in a human-derived IL-34cDNA into a codon tgt encoding cysteine; co-transfecting an Sf9 insect cell with a transfer plasmid containing an S98C IL-34 mutant encoding sequence and BacVector-3000 baculovirus DNA to prepare a baculovirus for expressing S98C IL-34; and then adopting the baculovirus for transducing an HEK 293 cell to express the S98C IL-34. Compared with native IL-34, the S98C IL-34 obtained by the method has the thermal stability improved. Through M-NFS-60 cell proliferation experiments and CSF-1R phosphorylation experiments, the activity of the S98C IL-34 is proved to have no significant difference with that of the native IL-34.

Description

technical field

[0001] The invention relates to a method for improving the thermal stability of interleukin IL-34 (interleukin-34, IL-34), in particular to a method for improving the thermal stability of IL-34 through amino acid site-directed mutation, which belongs to biological technology field. Background technique

[0002] Interleukin-34 (interleukin-34, IL-34) is a newly discovered cytokine (HaishanLin et al. Science 2008). It is a new ligand of colony-stimulating factorl receptor (CSF-1R), which can specifically recognize and activate CSF-1R independently. The biologically active form of IL-34 is a dimer. The molecular weight of the biological dimer is 50KD, and the molecular weight of each monomer is 25KD. The amino acid sequence of the monomer is:

[0003]

[0004]

[0005] Since IL-34 was identified as a new ligand of CSF-1R, its function and structure have been deeply studied. Although IL-34 has the effect of activating CSF-1R similar to CSF-1, but the act...

Examples

Embodiment 1

[0033] 1. Construction of expression plasmids

[0034] Material:

[0035] The cDNA of IL-34 was purchased from Origene Company. The cloning vector used was a BacMam vector.

technical field

[0001] The invention relates to a method for improving the thermal stability of interleukin IL-34 (interleukin-34, IL-34), in particular to a method for improving the thermal stability of IL-34 through amino acid site-directed mutation, which belongs to biological technology field. Background technique

[0002] Interleukin-34 (interleukin-34, IL-34) is a newly discovered cytokine (HaishanLin et al. Science 2008). It is a new ligand of colony-stimulating factorl receptor (CSF-1R), which can specifically recognize and activate CSF-1R independently. The biologically active form of IL-34 is a dimer. The molecular weight of the biological dimer is 50KD, and the molecular weight of each monomer is 25KD. The amino acid sequence of the monomer is:

[0003]

[0004]

[0005] Since IL-34 was identified as a new ligand of CSF-1R, its function and structure have been deeply studied. Although IL-34 has the effect of activating CSF-1R similar to CSF-1, but the act...

Examples

Embodiment 1

[0033] 1. Construction of expression plasmids

[0034] Material:

[0035] The cDNA of IL-34 was purchased from Origene Company. The cloning vector used was a BacMam vector.

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