A solid-state nuclear magnetic resonance method for the detection of weakly stable base pairs in RNA
A technology of weak stability and nuclear magnetic resonance, which is applied in the analysis of nuclear magnetic resonance, material analysis through resonance, and measurement devices, can solve the problem of lack of specific detection of weak stability base pairs, and achieve specificity Detection, overcoming difficult detection difficulties, avoiding the effects of dipolar coupling interactions
Active Publication Date: 2022-02-18
PEKING UNIV +1
View PDF6 Cites 0 Cited by
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Although traditional structural biology techniques, such as liquid NMR, X-ray crystal diffraction technology, and cryo-electron microscopy, can analyze the structure of RNA and obtain structural information about hydrogen bonds, it is difficult to detect the strength of hydrogen bonds in RNA, and The strength of base pair stability in RNA is still a difficulty in various technical means, especially the lack of experimental methods for specific detection of weakly stable base pairs (J.Sheng, et al., Nucleic Acids Res, 2013.)
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View moreImage
Smart Image Click on the blue labels to locate them in the text.
Smart ImageViewing Examples
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0049] 1. RNA sample preparation:
[0050] In this example, the aptamer part (riboA71) of the riboswitch RNA (add-riboswitch) regulating the add gene was used as a model to test the pulse sequence. The RNA is composed of 71 ribonucleotides, which is the aptamer region of add-riboswitch, and the ligand is adenine. Three kinds of riboA71-related samples were prepared in the present invention, namely the complex of riboA71 and ligand (denoted as riboA71-adenine), riboA71 without ligand (denoted as W-riboA71) and the mutation of G30 and U19 to A30 and A19 The riboA71-adenine complex (referred to as M-riboA71-adenine), the structure is as image 3 shown.
[0051] (1) Construction of DNA template
[0052] In this example, three kinds of riboA71 were obtained by in vitro transcription method. First, the corresponding DNA template needs to be designed according to the target RNA. The DNA template consists of two complementary single strands of DNA, the forward strand (R strand) an...
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More PUM
| Property | Measurement | Unit |
|---|---|---|
| purity | aaaaa | aaaaa |
Login to View More
Abstract
The invention discloses a solid-state nuclear magnetic resonance method for detecting weakly stable base pairs in RNA, and relates to the field of solid-state nuclear magnetic resonance spectroscopy. By designing a novel two-dimensional hydrogen detection solid-state nuclear magnetic resonance pulse sequence 2D WaterREXSY, the base pairs with weak stability in RNA are detected. In the pulse sequence, the Lee‑Goldburg spin-lock field on the proton was used to observe the chemical exchange of water with the exchangeable protons in RNA, and by suppressing 1 H spin diffusion and dipole coupling interact to achieve specific detection of chemical exchange processes. The exchange rate of the exchangeable proton in the weakly stable base pair and water is faster, and the signal is stronger, so as to realize the specific detection of the weakly stable base pair.
Description
technical field [0001] The invention relates to the field of solid-state nuclear magnetic resonance spectroscopy, in particular to a method for detecting weakly stable base pairs in RNA based on solid-state nuclear magnetic resonance technology and the chemical exchange between exchangeable protons and water in RNA. Background technique [0002] RNA is an important biological macromolecule, which plays an important role in transmitting genetic information, regulating gene expression, and resisting and treating diseases (Bandyra, K.; Luisi, B., RNA, 2015.). The biological functions of RNA are closely related to the diverse structures and complex dynamics of RNA (J. Chen and W. Zhang, J Chem Phys, 2012.). One of the foundations of RNA structure is base pairs stabilized by hydrogen bonds. The stability of the base pair is determined by the stability of the hydrogen bond forming the base pair, which is closely related to the type of base forming the base pair and the position o...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More Application Information
Patent Timeline
Login to View More Patent Type & Authority Patents(China)
IPC IPC(8): G01N24/08
CPCG01N24/08G01N24/087
Inventor 王申林赵莎温子扬邹梦冰张立新
Owner PEKING UNIV