S-Space College of Agriculture and Life Sciences (농업생명과학대학) Dept. of Food and Animal Biotechnology (식품·동물생명공학부) Theses (Master's Degree_식품·동물생명공학부)
Enhancing gene expression through stabilization of mRNA in Escherichia coli
- Issue Date
- 서울대학교 대학원
- E. coli; mRNA 안정성; Mrna; stem-loop structure; Stability; hairpin; Stem-loop structure; chaperone; Hairpin; heat shock protein; Chaperone; amylase; Heat shock protein; Amylase
- Thesis (master`s)--서울대학교 대학원 :농생명공학부,2004.
- Expression level of a gene can be modulated by changing stability of mRNA in Escherichia coli.
In order to develop an efficient E. coli expression system, the effects of secondary structures
on the 5-untranslated region (5''-UTR) of mRNA and heat shock proteins on stability of mRNA
were studied. The plasmids conferring different stem-loop structures on the 5''-end of mRNA
(pHY1, pHY2, pHY3, and pHY4) were constructed and the genes encoding amylase were tested. The
degree of stabilization was varied depending on the stem-loop structures. As an additional
stabilizing factor, coexpressed heat shock proteins DnaKJ, GrpE, GroELS encoded by pG-KJE6, or
GroELS encoded by pGro7, were involved in the stabilization of mRNA. The degree of stabilization
was varied depending on the stem-loop structures and variety of heat shock proteins. These
results suggest that secondary structures and heat shock proteins can enhance gene expression by
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