Conserved regional 3' grouping of rare codons in the coding sequence of ocular prosecretory mitogen lacritin.

ABSTRACT:

Purpose

Lacritin is a prosecretory mitogen in tears and, although a tear protein, it promotes basal tearing and lacrimal gland secretion. Since scale up is relevant to its potential use in the treatment of dry eye, we explored various mutagenic strategies to alter the stability, solubility, and translational efficiency of nascent lacritin, and discovered 3' clustering of rare human codons.

Methods

Site-directed mutagenesis of lacritin coding cDNA "pLAC" generated 24 different nonsynonymous and 13 synonymous mutations. Nonsynonymous mutations altered amino acids with nonpolar, basic or acidic side chains to serine. Synonymous mutation progressively optimized human codons that are rare or uncommon in Escherichia coli without changing the amino acid specified. These changes wer

SUBMITTER: McKown RL 

PROVIDER: S-EPMC3604909 | biostudies-literature | 2013 Mar

REPOSITORIES: biostudies-literature