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Fusion protein SUMO1-PLA2, and preparation method and medical use thereof

A SUMO1-PLA2, TF-SUMO1-PLA2 technology, applied in biochemical equipment and methods, chemical instruments and methods, hybrid peptides, etc., can solve the problem of low expression of PLA2 protein, inconvenient purification, reactogenicity of expression products and poor performance. Immunogenicity is not as good as soluble protein, etc., to overcome the effect of low protein expression

Inactive Publication Date: 2016-05-04
JILIN UNIV
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention further provides a method for preparing the fusion protein SUMO1-PLA2. The pColdTF cold-induced expression vector is used to efficiently prepare the fusion protein SUMO1-PLA2, which overcomes the low expression level of the PLA2 protein and the presence of the expression product in the form of inclusion bodies in the existing method. Inconvenience, and the reactogenicity and immunogenicity of the expressed product are not as good as soluble proteins.

Method used

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  • Fusion protein SUMO1-PLA2, and preparation method and medical use thereof
  • Fusion protein SUMO1-PLA2, and preparation method and medical use thereof
  • Fusion protein SUMO1-PLA2, and preparation method and medical use thereof

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Embodiment 1

[0043] Gene Amplification and Fusion

[0044] Acquisition of full-length fusion gene SUMO1-PLA2 according to the present invention

[0045] 1. To obtain the chicken SUMO1 gene, the steps are as follows:

[0046] Retrieve the complete sequence of the chicken SUMO1 gene. (From NCBI Nucleic Acid Database)

[0047] Chicken SUMO1 nucleic acid sequence

[0048] 1ATGTCGGACCAGGAAGCAAAGCCTTCAGCTGAGGACTTAGGAGATAAGAAAGAGGGGGAAT

[0049] ACATTAAACTCAAAGTCATTGGGCAGGACAGCAGTGAAATTCACTTCAAGGTGAAAATGAC

[0050] AACACACCCTCAAGAAACTCAAAGAATCATACTGTCAAAGACAGGGTGTTCCAATGAACTCACTCAGGTTCTCTTCGAGGGTCAGAGAATTACTGATAATCATACCCCCAAGGAGCTGGGGA

[0051] 288TGGAGGAGGAAGATGTGATTGAAGTTTATCAGGAACAGACCGGT

[0052] protein sequence

[0053] 1MSDQEAKPSAEDLGDKKEGEYIKLKVIGQDSSEIHFKVKMTTHLKKLKESYCQRQGVPMNS

[0054] 96LRLFEGQRITDNHTPKELGMEEEDVIEVYQEQTG

[0055] The chicken genome was extracted, and the SUMO1 gene sequence was obtained by PCR.

[0056] Upstream Primer 1:

[0057] CATATG ATGTCGGACCAGGAAGCAAA...

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Abstract

The invention provides a fusion protein SUMO1-PLA2, and a preparation method thereof. The invention also provides a medical use of the SUMO1 fusion protein to solve the technical problem of insoluble expression of PLA2 in Escherichia coli. The fusion protein SUMO1-PLA2 is efficiently prepared by using a pColdTF cold-induced expression vector. The defects of low PLA2 protein expression level, subsequent purifying inconvenience brought by existence of an expression product in an inclusion body form, and lower reactogenicity and immunogenicity of the expression product than soluble proteins in the prior art are overcome in the invention.

Description

technical field [0001] The invention provides a fusion protein SUMO1-PLA2 and a preparation method, and the invention also provides the medical application of the SUMO1 fusion protein, which belongs to the field of biotechnology. Background technique [0002] SUMO (Smallubiquitin-related modifier) ​​is a newly discovered protein post-translational modification widely present in eukaryotes and plays an important role in life activities. The role of SUMOylation in vivo includes regulating the interaction between proteins, regulating the mutual transport of proteins in the nucleus and the cytoplasm, and regulating the cell cycle. Since the substrate proteins modified by SUMOylation are some very important cell cycle regulatory proteins, such as p53, PML and RanGAP1, etc., the process of SUMOylation and de-SUMOylation plays a very important biological function in cells, and high Sensitive substrates and analytical methods are the key to related research. [0003] Phospholipase...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N9/18C12N15/70C12Q1/37
Inventor 艾永兴于佩峰邓晨吴山力郑海南王梦云赵程程郝林琳于浩张玉静
Owner JILIN UNIV
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