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Human dry cell factor macrophage clong irritant factor bifunction protein and its preparation

A technology of colony stimulating factor and bifunctional protein, which can be used in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., and can solve problems such as damage to various organs

Inactive Publication Date: 2004-12-22
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, long-term and high-dose use of M-CSF can inhibit the hematopoietic process and cause damage to various organs

Method used

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  • Human dry cell factor macrophage clong irritant factor bifunction protein and its preparation
  • Human dry cell factor macrophage clong irritant factor bifunction protein and its preparation
  • Human dry cell factor macrophage clong irritant factor bifunction protein and its preparation

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Experimental program
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Embodiment Construction

[0069] 1.1. Use pUC18-SCF as template, use primers 1 and 2 to synthesize SCF gene fragment by PCR method and clone it into pUC18 plasmid vector to obtain pUC18-5'SCF plasmid, use pUC18-M-CSF as template, use primer 3 , 4 synthesize the M-CSF gene fragment by PCR method, and clone it into pUC18 plasmid vector to obtain pUC18-3'M-CSF plasmid, and clone the SCF gene fragment into pUC18-3'M-CSF to obtain pUC18-SCF / M- CSF.

[0070] 1.2. The SCF / M-CSF gene fragment was digested from pUC18-SCF / M-CSF, and inserted between the Xba I and KpnI sites of the AcNPV transfer vector to obtain the recombinant transfer vector pVL1392-M-CSF / SCF. Sf9 was co-transfected with pVL1392-SCF / M-CSF and AcNPV DNA, and the recombinant virus AcNPV-SCF / M-CSF without wild-type virus was selected by screening and combined with TF-1 cell viability assay.

[0071] 2. Take the doubling time as 21 hours, live cells account for 90%, and the density is 1×10 6 Cells / mLSf9 cells were infected with the recombinant v...

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Abstract

A dual-function protein human stem cell factor-macrophage colony stimulating factor (hSCF / M-CSF) and a process for preparing its recombinant one (rhSCF / M-CSF) by efficient secretion expression in insect are disclosed. It can be used for rebuilding and restoring the hematopoietic and immune functions of a patient after radiotherapy, chemicotherapy and bone marrow transplantation.

Description

1. Technical field [0001] The invention belongs to the technical field of polypeptide medicine prepared by genetic engineering. 2. Background technology [0002] Stem cell factor (SCF, also called Kit ligand, Steel factor, mast cell growth factor, etc.) is a multifunctional cytokine that can promote the proliferation of early hematopoietic stem cells without inducing cell differentiation. Cytokines act synergistically to promote the survival and proliferation of hematopoietic stem / progenitor cells and various hematopoietic cells. Its receptors are widely distributed in different series and generations of hematopoietic cells, including erythrocytes, mononuclear giant cells, and megakaryocytes. SCF can enhance the proliferative ability of these cells and activate these cells to make them more sensitive to the stimulation of various hematopoietic cytokines. Clinically, it is mainly used in the reconstruction and recovery of hematopoietic and immune functions after radiotherap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/33C12N15/62C12P21/00
Inventor 秦浚川陈涛朱洁臧宇辉
Owner NANJING UNIV
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