Recombinant extracellular matrix-like proteins with repetitive elastin or collagen-like functional motifs.
Using overlap elongation PCR, we created repetitive DNA libraries encoding the elastin VPGVG and collagen-like GERGDRGDP sequences. From these libraries we isolated two repetitive DNA sequences, Col-5 encoding [(GERGDRGDP)(5)GER], and Ela-16 encoding [(VPGVG)(16)VPG]. Both proteins were expressed as thioredoxin fusion proteins. The resulting recombinant extracellular matrix-like proteins had the expected properties (cell adhesive ability and thermally responsive structural change) of the functional motif sequence unit used.
Kurihara
Hiroyuki
H
School of Engineering, The University of Tokyo, Bunkyo-ku, 113-8656 Tokyo, Japan.
Morita
Tetsuaki
T
Shinkai
Masashige
M
Nagamune
Teruyuki
T
D020816:Amino Acid Motifs; D000818:Animals; D001483:Base Sequence; D001709:Biotechnology / Q000379:methods*; D002448:Cell Adhesion; D003094:Collagen / Q000737:chemistry*; D004247:DNA / Q000737:chemistry; D004549:Elastin / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000378:metabolism*; D015723:Gene Library; D006358:Hot Temperature; D051379:Mice; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D009841:Oligonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry*; D017422:Sequence Analysis, DNA; D013696:Temperature; D013879:Thioredoxins / Q000737:chemistry
D009841:Oligonucleotides; D011994:Recombinant Proteins; D013879:Thioredoxins; D003094:Collagen; D004247:DNA; D004549:Elastin