Cloning and expression of a functional fucose-specific lectin from an orange peel mushroom, Aleuria aurantia.
Aleuria aurantia lectin (AAL) shows sugar-binding specificity for L-fucose. A lambda gt11 expression library was constructed from A. aurantia poly(A) RNA and screened with a polyclonal antiserum directed against AAL. An immunopositive clone carrying 1.3-kb EcoRI fragment was obtained. The fragment encoded AAL, but lacked a nucleotide sequence corresponding to the two amino-terminal amino acids. The 5'-terminal part of the fragment was replaced with a chemically synthesized DNA fragment and inserted into an expression vector to yield a plasmid pKA-1. Escherichia coli carrying pKA-1 expressed functional AAL and the recombinant AAL showed the same immunological properties as those of natural AAL.
Fukumori
F
F
Research Institute, Nichirei Co., Tokyo, Japan.
Takeuchi
N
N
Hagiwara
T
T
Ito
K
K
Kochibe
N
N
Kobata
A
A
Nagata
Y
Y
D000595:Amino Acid Sequence; D001203:Ascomycota / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D037102:Lectins / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D015183:Restriction Mapping
D037102:Lectins; C027112:fucose-binding lectin