Recombinant targeting fusion protein trail-sgrsa-gst and its anti-tumor application
A fusion protein, targeting technology, applied in anti-tumor drugs, recombinant DNA technology, drug combination and other directions, can solve problems such as interference with normal cell function
Inactive Publication Date: 2021-04-06
THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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Problems solved by technology
The selective effect of TRAIL on tumor cells makes people see the hope of targeted therapy, but TRAIL protein also has some very practical defects: ① Only a large dose of TRAIL protein can inhibit the proliferation of tumor cells in vivo , but large doses often have a significant killing effect on normal human liver cells, liver toxicity has become the main concern in the application of large doses of TRAIL; ②The body widely expresses TRAIL receptors Injection of exogenous TRAIL protein is bound to interfere with the function of normal cells
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[0029] specific implementation plan
[0030] Existing studies have shown that urokinase-type plasminogen activator (u-PA), an important tumor marker, is highly expressed in colon cancer. Based on the anti-tumor biological properties of TRAIL protein and the close relationship between uPA and tumors, Using the natural enzymatic activity of uPA on the surface of colon cancer cells, we designed a TRAIL-GST fusion protein expression plasmid with a uPA-specific cleavage site sequence ( figure 1 A). The GST tag is located at the C-terminal of the human TRAIL protein, and the u-PA specific cleavage site sequence is inserted between the TRAIL protein and the GST tag, named TRAIL-SGRSA-GST. The PCR primers used to amplify the human TRAIL DNA sequence are shown in Table 1, in which the primers contain sequences encoding u-PA-specific cleavage sites, and the primers used for amplification are named hTRAIL-F and hTRAIL-µPA-R. The partial sequence of human TRAIL was amplified with cDNA r...
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The present invention relates to the field of genetically engineered protein drugs, in particular to a recombinant targeting fusion protein TRAIL-SGRSA-GST and its anti-tumor application, wherein the recombinant targeting fusion protein TRAIL-SGRSA-GST is represented by SEQ ID NO.2 Comprising the amino acid sequence shown; the expression type recombinant comprising the gene, the engineering bacteria comprising the expression type recombinant, the recombinant targeting fusion protein TRAIL-SGRSA-GST expressed and isolated by the engineering bacteria, and the targeting fusion The specific killing effect of the protein on a class of colon cancer malignant tumor cells, the recombinant targeting fusion protein TRAIL-SGRSA-GST and its anti-tumor application of the technical solution Compared with the prototype TRAIL, the recombinant targeting fusion protein TRAIL-GST of the present invention SGRSA‑GST has a better application prospect.
Description
technical field [0001] The present invention relates to the field of genetically engineered protein drugs, more specifically, the present invention relates to a gene encoding a novel recombinant targeted anti-tumor drug fusion protein TRAIL-SGRSA-GST, an expression-type recombinant vector comprising the gene, an expression-type recombinant The engineering bacterium of the carrier, the targeting fusion protein TRAIL-SGRSA-GST expressed and isolated by the engineering bacteria and the specific killing effect of the targeting fusion protein on a kind of colon cancer malignant tumor cells. Background technique [0002] Colon cancer is one of the common malignant tumors, and its incidence rate ranks among the top three of various malignant tumors in western developed countries. In recent years, its incidence rate has a clear upward trend, especially in my country. Structural changes have significantly increased its incidence. Although great progress has been made in the pathogene...
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IPC IPC(8): C07K19/00C12N15/70C12N1/21A61K38/17A61K47/64A61P35/00
CPCA61K38/177A61K47/42A61P35/00C07K14/70575C07K2319/00C12N9/1088C12N9/6459C12N15/70C12Y205/01018C12Y304/21068
Inventor 马丹蔡振宇
Owner THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV