Method for efficient soluble expression of recombinant human interleukin-15

An interleukin, soluble technology, applied in the field of genetic engineering, can solve the problems of difficult purification, many purification steps, low expression, etc., and achieve the effect of improving natural activity, reducing endotoxin and low cost

Inactive Publication Date: 2012-07-11
无锡赛特基因生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] There are reports on the prokaryotic expression and purification of recombinant hIL-15 at home and abroad, but they are all expressed in inclusion bodies. The disadvantages are ① low expression, and the recombinant protein accounts for 5-10% of the total

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  • Method for efficient soluble expression of recombinant human interleukin-15
  • Method for efficient soluble expression of recombinant human interleukin-15
  • Method for efficient soluble expression of recombinant human interleukin-15

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Embodiment Construction

[0034] Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. Experimental methods not indicating specific conditions in the following examples are usually listed in molecular cloning according to conventional conditions: the conditions described in the laboratory manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer's suggestion condition.

[0035] 1. Construction of recombinant expression vector pQE30 / hIL-15

[0036] Firstly, optimize the preferred codons of Escherichia coli and mutate part of the human interleukin-15 coding sequence, and add BamHI and SalI restriction sites at its 3' and 5' ends, namely CGCGGATCC and ACGCGTCGAC. The sequence with restriction sites was synthesized for the whole gene, and then transferred into the pUC57 plasmid ...

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Abstract

The invention disclose a method for efficient soluble expression of recombinant human interleukin-15, which comprises an optimized coding sequence of recombinant human interleukin-15 and a recombinant expression vector pQE30/hIL-15, and an engineering cell that can express soluble recombinant human interleukin-15.

Description

technical field [0001] The invention relates to the field of genetic engineering. More specifically, the present invention provides an optimized coding sequence of recombinant human interleukin-15, the construction of the corresponding recombinant expression vector pQE30 / hIL-15, and a method for constructing engineered cells that highly express soluble recombinant human interleukin-15 . Background technique [0002] In 1994, Grabstein et al. first discovered a factor in the supernatant of monkey kidney epidermal cell line CV-1 / EBVA that can maintain the proliferation of IL-2-dependent cell line-T cell line (CTLL), and then the gene was successfully cloned and named as Interleukin-15 (interleukin 15, IL-15). The human interleukin-15 (hIL-15) gene is located at the 4q31 locus of the human chromosome. It is 32kb long, has 6 exons, and 7 introns. The cytokines encoded in the same region also include epidermal growth factor (EGF ), fibroblast growth factor (FGF) and IL-2 genes...

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Application Information

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IPC IPC(8): C12N15/24C12N1/21C12N15/70C12R1/19
Inventor 朱凌王柯周凡凡刘炜
Owner 无锡赛特基因生物科技有限公司
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