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MRNA stem ring structure probe, prepraring method thereof and application of the same in EMSA

A stem-loop structure and probe technology, applied in EMSA, mRNA stem-loop structure probe and its preparation field, can solve many problems such as time, achieve low price, facilitate quantitative analysis, and avoid laboratory safety. Effect

Inactive Publication Date: 2008-09-24
JIANGSU UNIV
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Problems solved by technology

The above-mentioned problems are more obvious in the case of RNA-protein interactions. Although the use of radioactive isotope-labeled probes has extremely high sensitivity, it still takes a lot of time to optimize the binding reaction conditions and gel electrophoresis. condition
[0016] In summary, the current study of mRNA-protein interaction is far less extensive than that of DNA-protein or nucleic acid hybridization, but due to the important regulatory role of this interaction at the level of cellular protein expression, the research potential is huge

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  • MRNA stem ring structure probe, prepraring method thereof and application of the same in EMSA
  • MRNA stem ring structure probe, prepraring method thereof and application of the same in EMSA
  • MRNA stem ring structure probe, prepraring method thereof and application of the same in EMSA

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Embodiment 1

[0051] For the research on mRNA-protein interaction, the earlier and more in-depth research is the interaction between iron-responsive elements (IRE) in mRNA and iron-regulatory protein (IRP) contained in the cytoplasm. effect. Studies have made it clear that the transferrin receptor and the IRE of the divalent cation transporter DMT1 are located at the 3' UTR, while ferritin (including L-type and H-type), δ-amino-γ-levulinic acid synthase (ALAS) and ferroportin1 are located at the 5' UTR. In iron deficiency, IRP binds to the IRE in these mRNAs, thereby enhancing the stability of the mRNAs of the transferrin receptor and the divalent cation transporter DMT1, and inhibiting the protein translation of the mRNAs of the ferritins ALAS and ferroportin1, whereas when iron When too much IRP dissociates from these IREs, the opposite effect occurs on mRNA. Thus, IRE-IRP interactions play critical regulatory roles in cellular iron uptake, iron storage, iron utilization, and iron relea...

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Abstract

The invention relates to the field of biological technology, in particular to a probe of mRNA stem-loop structure and a preparation method thereof as well as application on EMSA thereof. The probe of mRNA stem-loop structure is fabricated according to the following method: according to the sequence of nuclei acid of the objective stem-loop structure, chemical synthesis method is adopted to directly synthesize a probe with the sequence length of nuclei acid of no more than 70nt, then fluorescence mark method is adopted to mark the probe. Applying the probe of stem-loop structure with fluorescence mark into EMSA technology has the advantages of the high specificity, the high sensitivity, the repeatability and the stability, simultaneously the preparation method is simple and the price is low.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an mRNA stem-loop structure probe, a preparation method thereof, and its application in EMSA. Background technique [0002] Whether eukaryotic or prokaryotic, mRNA contains not only genetic information but also regulatory elements. It has been found that the regulatory elements are located in the 5' or 3' untranslated region (UTR), and the double-stranded structure formed by the local region is a "hairpin" structure, also known as a "stem-loop structure" (stem -loop). Different regulatory elements are involved in the regulation of different functions. Cells contain corresponding specific binding proteins, called RNA-binding proteins (RNA-binding proteins), which, after binding to the specific stem-loop structure on the target mRNA, affect the stability (half-life) of mRNA and the initiation or activation of protein translation. Processes such as peptide chain extension play an imp...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C07H21/02C12N15/10
Inventor 肖德生刘梅钱海
Owner JIANGSU UNIV
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