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STRAIL protein and primer thereof in crassostrea rivularis for resisting human tumor HL60

A near river oyster and protein technology, applied in the direction of animal/human protein, tumor necrosis factor, DNA/RNA fragments, etc., can solve the problems of low efficacy and side effects, few basic research data, low tumor cytotoxicity, etc., and achieve easy Good storage, safety, and high specificity

Active Publication Date: 2012-01-04
ZHEJIANG UNIV
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  • Summary
  • Abstract
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  • Claims
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Problems solved by technology

However, most of the above are mixtures of polysaccharides and other substances, which are immunoregulatory products, and there are few basic research data, and the toxicity to tumor cells is low, so little is known about their efficacy and side effects.

Method used

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Examples

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Embodiment Construction

[0012] The methods of molecular biology used in the following examples of the present invention are all known techniques.

[0013] In the present invention, the Oyster Oyster sTRAIL protein is the tumor necrosis factor-related apoptosis-inducing ligand soluble cytokine protein, referred to as CasTRAIL, wherein Ca represents Oyster Oyster Oysteria, and sTRAIL represents the tumor necrosis factor-related apoptosis-inducing ligand soluble cytokine; HL60 is the acute Promyelocytic leukemia cell lines.

[0014] Implementation steps of the present invention include:

[0015] 1. Using Oyster oyster cDNA as a template, design primers for amplifying the sTRAIL protein gene, and obtain the complete sequence of the gene encoding sTRAIL protein.

[0016] 2. Cloning the sTRAIL gene fragment into a suitable pMD19-T vector (purchased from Takara Company, Japan) using DNA recombination technology, and then digesting with restriction endonucleases, and pET-28(a) The prokaryotic expression ve...

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Abstract

The invention relates to the field of biotechnology, aiming at providing sTRAIL protein and a primer thereof in crassostrea rivularis for resisting human tumor HL60. The protein is provided with an amino acid sequence as shown in SEQ ID NO: 2; gene for encoding the protein is provided with an nucleotide sequence as shown in SEQ ID NO: 1. The invention is also applied to expand oligonucleotide primer of the gene of the protein as shown in a right claim 2. The sTRAIL protein product developed by gene engineering technique of the invention has the advantages of mass destruction to HL60 tumor cell, high specificity, good security, mass production and easy storage.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a tumor necrosis factor-related apoptosis-inducing ligand soluble cytokine protein and a primer thereof against human tumor acute promyelocytic leukemia cell line (HL60) in oyster near river. Background technique [0002] Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), also known as apoptin 2 ligand (APO2 ligand, APO-2L) (Wiley et al., 1995; Pitti et al. , 1996), under the action of metalloproteases, its extracellular domain can be degraded to form soluble cytokine sTRAIL (SolubleTRAIL). sTRAIL has the biological function of TRAIL (Bodmer et al., 2000). TRAIL is an important transmembrane cytokine in the TNF superfamily (Wiley et al., 1995; Pitti et al., 1996), which can selectively induce apoptosis in most human tumor cells: including brain, kidney, Tumor cells derived from liver, lung, stomach, colon, breast, prostate, pancreas, bile duct, cervix, skin and h...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/525C12N15/28C12N15/11
Inventor 吴信忠杨受保
Owner ZHEJIANG UNIV
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