Nucleic acid aptamer of aflatoxin B1 and application thereof

A technology of aflatoxin and nucleic acid aptamer, which is applied in the preparation of test samples, DNA/RNA fragments, recombinant DNA technology, etc., and can solve immunological detection such as cumbersome operation, high production cost, and restriction enzyme-linked immunosorbent assay Rapid technology development and other issues, to achieve high affinity and specificity, good repeatability and stability, and good application prospects

CN102517289AInactive Publication Date: 2012-06-27CHINA NAT ACAD NANOTECH & ENG
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2012-06-27
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to a nucleic acid aptamer which is in high-specificity and high-affinity combination with aflatoxin B1 and application thereof. The nucleic acid aptamer can be used for detection analysis and separating enrichment of aflatoxin B1, and has the advantages of being good in specificity, high in affinity, homogeneous in quality, good in stability, short in development cycle, low in production cost and convenient to use. The invention further relates to derivation sequences of a nucleic acid aptamer sequence and comprises sequences after modification.
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Description

(1) Technical field:

[0001] The invention belongs to the field of biotechnology, and relates to the design and preparation of an aflatoxin B1 nucleic acid aptamer combined with aflatoxin B1 with high specificity and high affinity by using molecular biology technology - SELEX technology and its application. (two) background technology:

[0002] Aflatoxin (AF) is a general term for a group of mycotoxins with similar chemical composition. It exists in the largest amount and has the strongest toxicity. The main producing bacteria are Aspergillus flavus and Aspergillus parasiticus, which are most commonly found in peanut products such as peanuts and grains and oils, corn, rice, cottonseed, some nut foods and feed, and dairy products. AF has strong toxicity, classified according to the toxicity level, which belongs to the super toxic level, and its toxicity is 10 times that of potassium cyanide and 68 times that of arsenic. The main target organ of its action is the liver. Acute ...

Examples

Embodiment

[0047] Embodiment: a kind of aflatoxin B1 nucleic acid aptamer, it is characterized in that it is nucleotide sequence

[0048] A DNA molecule represented by GCATCACTACAGTCATTACGCATCGGGTAATCCTAAAGCGGAACTGAGGAGTGGGAGGTAAATCGTGTGAAGTGCTGTCCC or its complementary nucleotide sequence.

[0049] In the present invention, the single-stranded DNA random library and primers used for SELEX are synthesized by Takara Company. The two ends are fixed sequences, and the middle is a random sequence of 35 bases: 5'GCATCACTACAGTCATTACGCATCG-(N35)-ATCGTGTGAAGTGCTGTCCC 3', the library capacity is 10 14 Above, primer 1: 5'GCATCACTACAGTCATTACGCA 3', primer 2: 5'GGGACAGCACTTCACACGAT 3', primer 3: 5'GCGTAATGACAAAAAAAAAAAACAT-C6-NH2 3', primer 4: 5'Biot in-GGGACAGCACTTCACACGAT 3'. Aflatoxin B1 was purchased from ALEXIS Company, magnetic beads were purchased from Invitrogen Company, oligonucleotide purification reagents were purchased from Qiagen Company, and PCR kits and T vectors were purchased from ...