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SELEX technology method taking clinical tumor pathological tissue section as target and application thereof

A technique of tissue sectioning and pathology, applied in the direction of recombinant DNA technology, biochemical equipment and methods, measurement/testing of microorganisms, etc., can solve problems such as the inability to simulate the cell growth environment, and achieve the effect of wide application value

Inactive Publication Date: 2010-01-06
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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Problems solved by technology

However, the application of established cell lines or isolated cells cultured in vitro still cannot simulate the cell growth environment in solid tumor tissues. , using the obtained specific aptamer to catch clinical and pathological tissue-specific markers will bring a new concept to the search for disease molecular markers, especially tumor-specific markers
At present, there is no SELEX technology method targeting clinical tumor pathological sections at home and abroad

Method used

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  • SELEX technology method taking clinical tumor pathological tissue section as target and application thereof
  • SELEX technology method taking clinical tumor pathological tissue section as target and application thereof
  • SELEX technology method taking clinical tumor pathological tissue section as target and application thereof

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Embodiment Construction

[0017] The present invention will be further described below by taking the paraffin-embedded tissue sections of clinical breast ductal carcinoma as the target, the subtractive SELEX screening technology with the normal tissue adjacent to the cancer as the subtractive target, and the identification of the obtained aptamer and the affinity capture of the target molecule.

[0018] 1. Synthesis of random single-stranded DNA oligonucleotide library and corresponding primers

[0019] GP30 library:

[0020] 5'-GCAATGGTACGGTACTTCC(30N)CAAAAGTGCACGCTACTTTGCTAA-3'

[0021] Primers:

[0022] Sense primer (Plong-1): 5'-GCAATGGTACGGTACTTCC-3'

[0023] Antisense primer (P11): 5'-TTAGCAAAGTAGCGTGCACTTTTG-3

[0024] Antisense primer (Pstem-loop):

[0025] 5'-GCTAAGCGGGTGGGACTTCCTAGTCCCACCCGCTTAGCAAAGTAGCGTGCACTTTTG-3'

[0026] Note: N represents any one of A, T, G, and C.

[0027] Plong-1 and P11 are used to amplify double-stranded products, and Plong-1 and Pstem-loop are used to amplif...

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Abstract

The invention discloses the establishment and application of clinical tumor pathological tissue section- SELEX technology method, belonging to the technical field of biology, relating to pathological tissue section reducing SELEX screening method and application thereof in preparation of disease molecule probe and screening of bio-marker. The invention obtains distinctive single chain DNA oligonucleotide genin of pathological tissue by establishing reducing SELEX screening method of paraffin embedding pathological tissue sections, and can be used as probe applied to detection, assistant diagnosis and the like of diseases such as tumor. The obtained oligonucleotide genin is amiable for purifying target molecules and used for separating and identifying molecule markers related to disease state, such as tumor marker.

Description

Field of invention: [0001] The present invention relates to the establishment and application of a subtractive SELEX screening method for clinical tumor pathological tissue slices. Through the subtractive SELEX screening of paraffin-embedded clinical tumor pathological tissue slices, a specific single-stranded DNA oligonucleotide ligand for identifying the pathological tissue is obtained. Specific diseases include tumor detection and auxiliary diagnosis. The obtained oligonucleotide ligands can also be used to identify molecular markers associated with disease states, such as tumor markers. Background technique: [0002] Different stages of disease occurrence and development will involve changes at various molecular levels. For example, the occurrence, development, and metastasis of tumors are related to changes in the structure, expression patterns, and expression levels of various genes. Theoretically, these large biomacromolecular markers, such as proteins and nucleic a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 邵宁生李少华丁红梅黄艳萍徐华曹晓晓杨光赵强沈倍奋
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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