Degenerate codon mixing for PCR-based manipulation of highly repetitive sequences.

ABSTRACT:

Objective

Repeat expansion of polyglutamine tracks leads to a group of inherited human neurodegenerative disorders. Studying such repetitive sequences is required to gain insight into the pathophysiology of these diseases. PCR-based manipulation of repetitive sequences, however, is challenging due to the absence of unique primer binding sites or the generation of non-specific products.

Results

We have utilised the degeneracy of the genetic code to generate a polyglutamine sequence with low repeat similarity. This strategy allowed us to use conventional PCR to generate multiple constructs with approximately defined numbers of glutamine repeats. We then used these constructs to measure the in vivo variation in autophagic degradation activity related to the different numbers of

SUBMITTER: Ratnayake D 

PROVIDER: S-EPMC5870680 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature