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MiRNA target gene double-binding-site fluorescent vector and construction method and application thereof

A dual-binding site, target gene technology, applied in the field of genetic engineering, can solve problems affecting life processes and other issues, and achieve the effects of meeting experimental needs, rapid detection, and shortening the experimental cycle

Pending Publication Date: 2019-06-25
湖南省畜牧兽医研究所
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AI Technical Summary

Problems solved by technology

In animals, it is usually the "seed sequence" (2-8 nucleotides) of miRNA that binds to the target mRNA, resulting in translational inhibition of the target mRNA, which in turn affects various life processes

Method used

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  • MiRNA target gene double-binding-site fluorescent vector and construction method and application thereof
  • MiRNA target gene double-binding-site fluorescent vector and construction method and application thereof
  • MiRNA target gene double-binding-site fluorescent vector and construction method and application thereof

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Embodiment Construction

[0058] The present invention provides a miRNA target gene double binding site fluorescent vector, which has two wild-type site fragments that bind to miR-135a-5p "seed sequence" respectively, the first mutation site and the second Fragments between wild-type sites, fragments between the first wild-type site and the second mutation site, and fragments between two mutation sites. It should be noted that the fragment between the first mutation site and the second wild-type site not only includes the sequence from the first mutation site to the second wild-type site, but also includes the first mutation The partial sequence upstream of the site and the partial sequence downstream of the second wild-type site; the fragment between the first wild-type site and the second mutation site not only includes the first wild-type site to the second The sequence between the mutation sites also includes the partial sequence upstream of the first wild-type site and the partial sequence downstre...

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Abstract

The invention relates to a miRNA target gene double-binding-site fluorescent carrier and a construction method and application thereof. The miRNA target gene double-binding-site fluorescent carrier comprises two wild-type site fragments bound with a miR-135a-5p seed sequence, a fragment between the first and second wild-type sites, a fragment between first and second wild-type sites, and a fragment between two mutation sites. The invention provides a miRNA target gene double-binding-site fluorescent vector, a construction method and an application thereof. The miRNA target gene double-binding-site fluorescent vector can be used for rapidly and directly detecting the combination of miRNA and a predicted target gene, save the research cost and shorten an experiment period.

Description

Technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a fluorescent vector and a construction method, in particular to a miRNA target gene double binding site fluorescent vector and a construction method and application thereof. Background technique [0002] With the rapid development of molecular biology research, gene-directed mutagenesis has become an important experimental technique and has been widely used in the study of gene expression and the relationship between protein structure and function. At present, PCR-mediated site-directed mutagenesis technology, that is, overlapping primer extension PCR, is the most common method of site-directed mutagenesis, which can be mutated at any position in the DNA segment. [0003] MicroRNA (miRNA) is a type of endogenous, conserved, non-coding small RNA with a length of 19-23 nucleotides. In 1993, scientists discovered for the first time in C. elegans a miRNA that regulates its devel...

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Application Information

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IPC IPC(8): C12N15/85C12N15/65C12Q1/6818
Inventor 陈晨崔清明罗璇谢菊兰张星邓缘刘莹莹朱吉任慧波胡雄贵左剑波彭英林
Owner 湖南省畜牧兽医研究所
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