Expression of redesigned mussel silk-like protein in <Emphasis Type="Italic">Escherichia coli</Emphasis> |
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Authors: | Yun Jung Yang Yoo Seong Choi Dooyup Jung Hyung Joon Cha |
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Affiliation: | (1) Biological Systems Department, Idaho National Laboratory, Idaho Falls, Idaho 83415, USA |
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Abstract: | Silks have been used widely for human beings due to their several extraordinary properties. Until now, the studies on silk
proteins have mainly focused on spiders and silkworms. Because silk properties are organism-dependent, novel silk protein
types can be found and developed through investigation of new silk-bearing organisms. We noticed that marine mussel has silk-like
domains containing many repeats with abundance of glycine and alanine. In the present work, we redesigned mussel-derived silk-like
gene sequence which contains alternating repeated and nonrepeated regions with optimized codons for Escherichia coli. For successful expression of recombinant mussel silk-like protein in E. coli cells, we employed several experimental strategies, including use of strong promoter, cold shock expression, and genetic
fusions. We observed significant repression on cell growths by even low expression levels of soluble mussel silk-like proteins
in cold shock- and glutathione s-transferase (GST) fusion-based systems. Thus, we finally used baculoviral polyhedrin protein
as a fusion partner and successfully expressed insoluble mussel silk-like protein with relatively high expression level and
without cell growth repression in E. coli. |
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