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Modified nanopores, compositions comprising same and applications thereof

A nanopore and cytolysin technology, applied in biochemical equipment and methods, microbial measurement/inspection, resistance to vector-borne diseases, etc.

Active Publication Date: 2022-05-31
鲁汶大学研究与发展中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, translocation of double-stranded or single-stranded DNA through nanopores with negatively charged amino acids towards the inner surface is ineffective

Method used

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  • Modified nanopores, compositions comprising same and applications thereof
  • Modified nanopores, compositions comprising same and applications thereof
  • Modified nanopores, compositions comprising same and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

Acids Res. (2004) 32: W665‑W667; and Dolinsky et al. Nucleic Acid Res. (2007) 35: W522‑

The Adaptive Poisson-Boltzmann Solver (APBS) described in W525) calculates the potential distribution of two pores in 150 mM NaCl

(Fig. 11, version B). In ClyA‑AS, the potential at the center of the pore is found to move from the cis inlet through the middle part and to the trans

mode inlets are increasingly negative (-2.6, -4.8 and -15.2mV on average, respectively). In the case of ClyA‑RR, it can be observed that in the pores

The rise in potentials at the cis-entrance and mid-portion of , averaged ‑0.3 and ‑1.1 mV, respectively. The potential at the transconstriction is similar to

almost dropped further to an average of ‑17.3mV. It should be noted that these values ​​are calculated when no external bias is applied.

[0222] Table 1: Electrical properties of the designed ClyA nanopore variants. By adding ~0.1ng of oligomeric protein to the cis side

cavity to test the activity of...

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Abstract

Provided herein relate to modified or mutated forms of cytolysin A (ClyA) and compositions comprising the same. In particular, modified or mutated forms of ClyA allow efficient capture and / or transfer of negatively charged target molecules or polymers through the modified or mutated ClyA nanopore at low or physiological ionic strength. Accordingly, also provided are methods of using modified or mutated forms of ClyA and compositions, eg, for characterizing negatively charged target analytes, eg, target polynucleotides.

Description

Modified nanopores, compositions containing the same and uses thereof CROSS-REFERENCE TO RELATED APPLICATIONS This application requires U.S. Provisional Application No. 62 / of submission on December 8, 2015 according to 35 U.S.C. § 119 (e) 264,709, the contents of which are incorporated herein by reference in their entirety. technical field [0003] Provided herein are modified or mutated forms of cytolysin A (ClyA) and compositions comprising the same. also provides For example, to characterize a target analyte such as a target polynucleotide using the modified or mutated form of ClyA and the composition Methods. Background technique [0004] Transmembrane pores, such as nanopores, have been used to identify small molecules or folded proteins and monitor chemical or folded proteins at the single-molecule level. enzymatic reaction. Electrophoretic translocation of DNA through nanopores reconstituted in artificial membranes in practical applications such as DNA sequ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6869
CPCC12Q1/6869G01N33/48721C12Q2565/631C07K14/001
Inventor 乔瓦尼·马利亚洛伦佐·弗兰切斯基尼蒂内·邦斯安德鲁·约翰·赫伦拉科马·尼尚萨·贾亚辛格伊丽莎白·杰恩·华莱士
Owner 鲁汶大学研究与发展中心
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