Method for obtaining miRNA (Ribose Nucleic Acid) candidate target gene and special reverse transcription primer for method

Abstract

The invention discloses a method for obtaining a miRNA (Ribose Nucleic Acid) candidate target gene and a special reverse transcription primer for the method. The method comprises the following steps of: (1), acquiring cDNA (Deoxyribonucleic Acid) first chains corresponding to mRNA which is complementary to a target miRNA seed sequence to obtain a storeroom A by using the DNA with a stem-loop structure and the target miRNA seed sequence as a reverse transcription primer; (2), acquiring cNDA first chains corresponding to mRNA of the eukaryote target gene tissue to obtain a storeroom B by using the DNA with a special joint and 14-20 deoxidized thymine nucleotides as a reverse transcription primer, wherein the structure of the reverse transcription primer B is the special joint-T14-20; and (3), synthesizing the storeroom A and the storeroom B to obtain a double-chain cDNA, then hybridizing to obtain a hybrid DNA molecule, amplifying the DNA segment between the target miRNA seed sequence and the special joint on the hybrid DNA module and obtaining the target miRNA candidate target gene according to the amplification products.

Description

technical field [0001] The invention relates to a method for obtaining miRNA candidate target genes and special reverse transcription primers thereof. Background technique [0002] MicroRNA (microRNA, miRNA) is a class of small molecule non-coding RNA, about 22 bases in size. It causes the degradation of target gene mRNA or the inhibition of target protein translation by binding to the 3'untranslated region (3'UTR) of the target gene. Studies have shown that the biological function of miRNA is realized by regulating the target gene, and miRNA participates in various biological processes such as cell proliferation, apoptosis, differentiation, metabolism, development, and tumor metastasis by acting on the corresponding target mRNA. Controls the formation, development, metabolism, and occurrence of certain diseases including animal and plant organs. [0003] miRNAs combine with target genes in an incomplete pairing mode, which makes the regulatory network of miRNAs and their ...

AI Technical Summary

Problems solved by technology

The data obtained by the biological experiment method is relatively accurate, but its disadvantage is that only one target gene can be proved at a time, and the throughput is not enough

Although the second version (V2.0) of the sheep genome has been released, the fourth version has not yet been published. See the official sequencing website ( http: / / www.animalgenome.org / sheep / ), because the version is low, the sequencing quality is not high, there are many untested fragments (gap), the genome data is not complete, and the 3'UTR sequence of a large number of genes is unknown

Therefore, it is difficult to obtain complete information on the regulatory network of sheep skeletal muscle miRNAs and their target genes using the above two methods.

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