Recombinant plasmid with human thymosin Beta-4gene

A technology of recombining plasmids and thymosin, which is used in gene therapy, genetic engineering, plant gene improvement, etc.

Active Publication Date: 2008-03-05
BEIJING NORTHLAND BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, the vectors used to carry Tβ4 as gene therapy mainly include adenovirus vector (AV) and adeno-associated virus vector (AAV). The infection rate is very high, and it is easy to carry genes into human cells. However, it is precisely because of their viral characteristics that the safety of gene therapy has become a concern.

Method used

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  • Recombinant plasmid with human thymosin Beta-4gene
  • Recombinant plasmid with human thymosin Beta-4gene
  • Recombinant plasmid with human thymosin Beta-4gene

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Experimental program
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Embodiment Construction

[0014] The following examples are only for illustration of the present invention, and do not cause any limitation to the present invention.

[0015] One), construction of naked plasmid pNL-Tβ4

[0016] First, the whole gene sequence of thymosin β4 is amplified by whole gene synthesis:

[0017] GCT AGC ATG TCT GGT GAC AAA CCC GAT ATG GCT GAG ATC GAG AAA TTC GAT

[0018] AAG TCG AAA CTG AAG AAG ACA GAG ACG CAA GAG AAA AAT CCA CTG CCT TCC

[0019] AAA GAA ACG ATT GAA CAG GAG AAG CAA GCA GGC GAA TCG TAA CTC GAG

[0020] The target fragments were digested with NdeI and XhoI to recover small fragments respectively, and the plasmid pNL was digested with XhoI and EcoR I to recover large fragments, and the three target gene fragments were ligated in T4 ligase system at 18°C. The recombinant plasmid was named pNL-Tβ4 (see Figure 1 for the specific structure). Then transform Escherichia coli DH5a with the CaCl2 method, spread on the LB plate containing 50ug / ml kanamycin, select the p...

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Abstract

A recombinant plasmid with human extrasin beta4-gene carrier is prepared by inserting human extrasin beta4-gene into eukaryote expression starter to construct recombinant plasmid, entering it into striped muscle cell, synthesizing T beta-4 by cell protein synthetic system and secreting to extracellular region. It can improve wound healed, prevent cell from death and be used to repair skin and heart damages.

Description

1. Technical field [0001] The invention belongs to the field of gene therapy, specifically, it is a recombinant plasmid constructed by inserting human thymosin β4 gene into a eukaryotic expression promoter. The plasmid can enter striated muscle cells, and use the protein synthesis system of the cell itself to synthesize Tβ4 in the cell, and secrete it out of the cell, so as to accelerate wound healing and prevent cell death. 2. Background technology [0002] The immune system is the body's defense system. The main cells involved in the immune response are T lymphocytes, B lymphocytes and macrophages. The thymus is the central immune organ for the development and differentiation of T lymphocytes. Thymus factors (or hormones) secreted by the thymus are a series of essential substances required for the development and differentiation of T lymphocytes, among which thymosin β4 (Tβ4) is a major thymus factor whose structure and function have been clarified. Thymosin-beta 4 enha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00C12N15/85C12N15/16C07K14/575A61K38/22A61P17/02A61P9/00
Inventor 马素永聂李亚许松山文美玉
Owner BEIJING NORTHLAND BIOTECH
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