Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Insensitive Nuclei Enhanced by Polarization Transfer (INEPT)01:15

Insensitive Nuclei Enhanced by Polarization Transfer (INEPT)

1.2K
Insensitive Nuclei Enhanced by Polarization Transfer (INEPT) is an advanced Nuclear Magnetic Resonance (NMR) technique specifically designed to detect and enhance the signals of low-abundance nuclei, such as carbon-13 and nitrogen-15, in small molecules. The fundamental principle behind INEPT is the transfer of polarization from a more abundant and highly polarizable nucleus, typically hydrogen-1, to the low-abundance nucleus of interest. This process effectively boosts the NMR signal of the...
1.2K
Interpreting ¹H NMR Signal Splitting: The (n + 1) Rule01:10

Interpreting ¹H NMR Signal Splitting: The (n + 1) Rule

2.9K
In the AX proton spin system, proton A can sense the two spin states of a coupled proton X, resulting in a doublet NMR signal with two peaks of equal (1:1) intensity. When proton A is coupled to two equivalent protons (AX2 spin system), the spin states of each X can be aligned with or against the external field, creating three possible scenarios. This results in a 1:2:1  triplet signal, where the central peak corresponds to the chemical shift of A and is twice as large or intense as the...
2.9K
¹³C NMR: ¹H–¹³C Decoupling01:04

¹³C NMR: ¹H–¹³C Decoupling

1.7K
The probability of having two carbon-13 atoms next to each other is negligible because of the low natural abundance of carbon-13. Consequently, peak splitting due to carbon-carbon spin-spin coupling is not observed in spectra. However, protons up to three sigma bonds away split the carbon signal according to the n+1 rule, resulting in complicated spectra.
A broadband decoupling technique is used to simplify these complex, sometimes overlapping, signals. Broadband decoupling relies on a...
1.7K
Double Resonance Techniques: Overview01:12

Double Resonance Techniques: Overview

870
Double resonance techniques in Nuclear Magnetic Resonance (NMR) spectroscopy involve the simultaneous application of two different frequencies or radiofrequency pulses to manipulate and observe two distinct nuclear spins. One important application of double resonance is spin decoupling, which selectively suppresses coupling with one type of nucleus while observing the NMR signal from another nucleus, simplifying the spectrum and enhancing resolution.
Spin decoupling is usually achieved by...
870

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Predominant HLA class I associations with lupus nephritis susceptibility, histology class and reduced renal function in patients with systemic lupus erythematosus: a retrospective case-control study.

Rheumatology international·2026
Same author

Healthcare utilization and costs in patients with rheumatoid arthritis using biologics or tofacitinib: a nationwide, population-based study.

Clinical rheumatology·2026
Same author

Longitudinal Trajectories and Predictors of Health-Related Quality of Life in Systemic Lupus Erythematosus: A 2-Year Longitudinal Cohort Study.

Rheumatology and therapy·2026
Same author

Genetic polymorphisms associated with therapeutic response and adverse effects to methotrexate in Taiwanese patients with rheumatoid arthritis: a real-world, hospital-based study.

Scientific reports·2026
Same author

A culturally adapted online Mediterranean diet intervention for metabolic dysfunction-associated steatotic liver disease (TIMA): study protocol for a randomized controlled trial.

Trials·2026
Same author

Three-metal-ion catalysis by ribonuclease P holoenzyme: a new mechanistic insight into transfer RNA maturation.

Research square·2026
Same journal

Expression of a recombinant DIVA antigen for differential diagnosis of H7N9 subtype avian influenza virus infected and vaccinated chickens.

Protein expression and purification·2026
Same journal

Prokaryotic expression, purification of Tldi1 protein and preparation of its polyclonal antibody in Salmonella Typhimurium.

Protein expression and purification·2026
Same journal

Soluble expression, two-step purification and tagmentation-compatible activity assessment of hyperactive Tn5 transposase using a GB1 fusion tag strategy.

Protein expression and purification·2026
Same journal

High-level soluble production of firefly luciferase in E. coli via promoter engineering.

Protein expression and purification·2026
Same journal

Rapid generation of Drosophila Schneider 2 (S2) cell lines and FACS-based isolation of high-yield soluble or membrane proteins.

Protein expression and purification·2026
Same journal

Heterologous expression and characterization of multi-resistant manganese superoxide dismutase from Thermus aquaticus in Escherichiacoli.

Protein expression and purification·2026
See all related articles

Related Experiment Video

Updated: May 1, 2026

Assembly and Purification of Prototype Foamy Virus Intasomes
10:20

Assembly and Purification of Prototype Foamy Virus Intasomes

Published on: March 19, 2018

5.2K

A streamlined method for preparing split intein for NMR study.

Yi-Zong Lee1, Yun-Tzai Lee1, Yi-Jan Lin2

  • 1Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu, Taiwan.

Protein Expression and Purification
|April 23, 2014
PubMed
Summary
This summary is machine-generated.

Researchers developed a streamlined method for producing split inteins, crucial for protein trans-splicing studies. This technique utilizes circular permutation to improve purification and enable structural analysis of split intein fragments.

Keywords:
Circular permutationInteinNMRProtein trans-splicingSplit intein

More Related Videos

NMR-Based Fragment Screening in a Minimum Sample but Maximum Automation Mode
09:19

NMR-Based Fragment Screening in a Minimum Sample but Maximum Automation Mode

Published on: June 4, 2021

3.9K
Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage
06:51

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

Published on: May 6, 2020

3.7K

Related Experiment Videos

Last Updated: May 1, 2026

Assembly and Purification of Prototype Foamy Virus Intasomes
10:20

Assembly and Purification of Prototype Foamy Virus Intasomes

Published on: March 19, 2018

5.2K
NMR-Based Fragment Screening in a Minimum Sample but Maximum Automation Mode
09:19

NMR-Based Fragment Screening in a Minimum Sample but Maximum Automation Mode

Published on: June 4, 2021

3.9K
Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage
06:51

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

Published on: May 6, 2020

3.7K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • Inteins are protein catalysts mediating protein splicing, essential for various biological processes.
  • Split inteins, comprising two complementary fragments, enable protein trans-splicing but are challenging to produce and purify.
  • Aggregation and concentration control issues hinder conventional split intein production for structural studies.

Purpose of the Study:

  • To develop an efficient and streamlined method for producing high-purity split inteins.
  • To overcome aggregation and concentration control challenges in split intein preparation.
  • To facilitate structural studies of split inteins and their association mechanisms.

Main Methods:

  • Incorporation of a circular permutation strategy into the Nostoc punctiforme DnaE intein (NpuInt).
  • Relocation of the backbone opening to the native split site via circular permutation.
  • Self-cleavage of expressed NpuInt into two complementary fragments.

Main Results:

  • A streamlined method for producing split NpuInt was established using circular permutation.
  • The circular permutation strategy preserves protein splicing activity, leading to immediate self-cleavage.
  • The tight association of split NpuInt fragments allows for efficient one-step purification.
  • NMR spectra were used to assign backbone and side chain resonances for the native split NpuInt.

Conclusions:

  • Circular permutation offers a simple and broadly applicable strategy for streamlined split intein production.
  • This method overcomes key limitations of conventional split intein preparation, enabling easier purification.
  • The developed technique facilitates structural and mechanistic studies of split inteins and protein trans-splicing.