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

13.9K Views

12:49 min

September 4th, 2011

September 4th, 2011

13,926 Views

0:05

Title

2:03

Site-directed Mutagenesis of Highly GC-rich KISSR Gene Sequence

3:48

Cell Treatment and Lysis

6:24

Immunoprecipitation of MycKISS1R

7:16

Western Blot Detection of MycKISS1R Immunocomplexes

8:45

Imaging and Quantification of MycKISS1R

10:10

Analysis of KISS1R

11:46

Conclusion

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.

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