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Nucleic acid aptamer specifically combined with KDR and application of nucleic acid aptamer

A nucleic acid aptamer and specific technology, applied in the field of molecular biomedical technology and medical biology, can solve the problems of time-consuming and labor-intensive, and achieve the effect of simple preparation method

Pending Publication Date: 2021-08-24
YANGZHOU UNIV
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Problems solved by technology

[0005] At present, there are no reports on KDR nucleic acid aptamers and their screening methods in scientific research, and the traditional SELE X screening process is time-consuming and labor-intensive. Only after multiple rounds of screening can aptamers with strong specificity be obtained. Therefore, it is urgent to establish a KDR nucleic acid aptamers that can specifically target, and need to provide a simple, convenient and effective screening method

Method used

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  • Nucleic acid aptamer specifically combined with KDR and application of nucleic acid aptamer
  • Nucleic acid aptamer specifically combined with KDR and application of nucleic acid aptamer
  • Nucleic acid aptamer specifically combined with KDR and application of nucleic acid aptamer

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Embodiment Construction

[0027] 1. Synthesize the original random ssDNA library and primers shown in the following sequences:

[0028] Raw random ssDNA library:

[0029] 5'-TGCGCTCTAGAGACGAAT(N)45CCTCAGCTGCAGGTATAC-3';

[0030] Upstream primer: 5'-TGCGCTCTAGAGACGAAT-3' (SEQ ID NO.2);

[0031] Downstream primer: 5'-GTATACCTGCAGCTGAGG-3' (SEQ ID NO.3);

[0032] Biotin-labeled downstream primer: 5'-Biotin-GTATACCTGCAGCTGAGG-3' (SEQ ID NO.4).

[0033] 2. Establish a screening method for nucleic acid aptamers based on CELL-SELEX

[0034] 1) Used in the whole screening process: the composition of binding buffer (BB) is 0.01M pH 7.4PBS, 250mM glucose, 5MM MgCl 2 .6H 2 O, 0.1mg / mL tRNA, 1mg / mL bovine serum albumin; the composition of wash buffer (WB) is 0.01M pH7.4PBS, 250mM glucose, 5MM MgCl 2 .6H 2 O.

[0035] 2) Establish a Cell-SELEX-based nucleic acid aptamer screening method

[0036] Through the KDR expressed on the cell surface, a cell-based nucleic acid aptamer screening method was establishe...

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Abstract

The invention belongs to the field of molecular biomedical technology and medical biology, and relates to a nucleic acid aptamer specifically combined with KDR, and screening, identification and application thereof. According to the KDR nucleic acid aptamer screening method based on the combination of the CELL-SELEX technology and the polyacrylamide gel electrophoresis, a nucleic acid aptamer specifically combined with KDR is screened out. The preparation method is simple and convenient, and the nucleic acid aptamer can carry molecular markers or chemotherapeutic drugs to be used for diagnosis and treatment of tumors (gastric cancer).

Description

technical field [0001] The invention belongs to the fields of molecular biomedical technology and medical biology, and relates to a nucleic acid aptamer specifically binding to KDR and its screening, identification and application, and specifically relates to the design, preparation, verification, and application of the nucleic acid aptamer in basic research and clinical practice. All aspects of treatment. Background technique [0002] Nucleic acid aptamer (also known as aptamer, aptamer) is a small piece of ssDNA or RNA that can specifically bind to a target molecule and is screened by the systematic evolution of ligands by exponential enrichment (SELEX) technology. . Aptamers have the advantages of a wide range of target molecules, strong affinity, and easy modification, and are widely used in molecular chemistry, food safety, clinical diagnosis, and treatment. [0003] Cell-SELEX is a new technology that uses the principle of combinatorial chemistry to construct a synth...

Claims

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Application Information

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IPC IPC(8): C12N15/115C12N15/10C12N15/11G01N33/574A61K31/7088A61P35/00
CPCC12N15/115C12N15/1048G01N33/57446G01N33/57484A61K31/7088A61P35/00C12N2310/16
Inventor 孔桂美黄瑶张小荣曾凤叶桐蔡凌宇王孝通
Owner YANGZHOU UNIV
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