Journal
/
Biology
/
Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders
JoVE Journal
Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biology
Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders

Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders

DOI:
10.3791/2897-v

•

12:49 min

•

September 04, 2011

•

, ,
1Division of Endocrinology and Metabolism, Department of Medicine,University of Miami Miller School of Medicine, 2Department of Molecular and Cellular Pharmacology,University of Miami Miller School of Medicine

Chapters

  • 00:05Title
  • 02:03Site-directed Mutagenesis of Highly GC-rich KISSR Gene Sequence
  • 03:48Cell Treatment and Lysis
  • 06:24Immunoprecipitation of MycKISS1R
  • 07:16Western Blot Detection of MycKISS1R Immunocomplexes
  • 08:45Imaging and Quantification of MycKISS1R
  • 10:10Analysis of KISS1R
  • 11:46Conclusion

Summary

Automatic Translation

Automatic Translation

Mutations in the kisspeptin receptor (KISS1R) are associated with reproductive disorders in patients. Here we describe how to introduce mutations of interest in the GC-rich sequence of KISS1R as well as the use of KISS1R constructs to characterize the degradation pathway of the receptor by immunoprecipitation and western blot.

Tags

MutagenesisGenetic MutationsGC-richKISS1 ReceptorReproductive DisordersKisspeptin ReceptorInactivating MutationsHypogonadotropic HypogonadismPrecocious PubertyFunctional StudiesRegulation Of ReproductionKISS1R GenePCR AmplificationPCR Enhancer SolutionDMSOExpression Vector

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/2897?article_type=v

Primary Culture and Plasmid Electroporation of the Murine Organ of Corti.
check_url/2897?article_type=v

Mutagenesis and Functional Analysis of Ion Channels Heterologously Expressed in Mammalian Cells
check_url/2897?article_type=v

Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides
check_url/2897?article_type=v

Rapid Genetic Analysis of Epithelial-Mesenchymal Signaling During Hair Regeneration
check_url/2897?article_type=v

Alternative Cultures for Human Pluripotent Stem Cell Production, Maintenance, and Genetic Analysis
check_url/2897?article_type=v

High-resolution Time-lapse Imaging and Automated Analysis of Microtubule Dynamics in Living Human Umbilical Vein Endothelial Cells
check_url/2897?article_type=v

Genetic and Biochemical Approaches for In Vivo and In Vitro Assessment of Protein Oligomerization: The Ryanodine Receptor Case Study
check_url/2897?article_type=v

Open-source Single-particle Analysis for Super-resolution Microscopy with VirusMapper
check_url/2897?article_type=v

Visualizing Genetic Variants, Short Targets, and Point Mutations in the Morphological Tissue Context with an RNA In Situ Hybridization Assay
check_url/2897?article_type=v

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing

Read Article