Expression and secretion of bovine beta-lactoglobulin in Saccharomyces cerevisiae.
A bovine beta-lactoglobulin (beta-LG) was expressed in Saccharomyces cerevisiae carrying bovine pre-beta-LG cDNA and secreted into its growth medium. The expression plasmid was constructed by inserting the whole coding region of the cDNA encoding pre-beta-LG between the promoter and terminator of the yeast glyceraldehyde 3-phosphate dehydrogenase gene of pYG100, a yeast expression vector. In the supernatant of the yeast growth medium, beta-LG with a native conformation was detected by sandwich ELISA, and its amount was estimated to be 1.1 mg/l. A Western-immunoblotting analysis revealed that beta-LG secrected in the growth medium had the same mobility as that of authentic bovine beta-LG. The N-terminal sequence was also identical with that of authentic mature bovine beta-LG.
Totsuka
M
M
Department of Agricultural Chemistry, Faculty of Agriculture, University of Tokyo, Japan.
Katakura
Y
Y
Shimizu
M
M
Kumagai
I
I
Miura
K
K
Kaminogawa
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002417:Cattle; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D015723:Gene Library; D007782:Lactoglobulins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D008321:Mammary Glands, Animal / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D011994:Recombinant Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
D004271:DNA, Fungal; D007782:Lactoglobulins; D011994:Recombinant Proteins