Expression and Secretion of Bovine β-Lactoglobulin in Saccharomyces cerevisiae |
| |
| Authors: | Mamoru Totsuka Yoshinori Katakura Makoto Shimizu Izumi Kumagai Kin-ichiro Miura Shuichi Kaminogawa |
| |
| Institution: | 1. Department of Agricultural Chemistry, Faculty of Agriculture;2. Department of Industrial Chemistry, Faculty of Engineering, The University of Tokyo, Bunkyo-ku, Tokyo 113, Japan |
| |
| Abstract: | A bovine β-lactoglobulin (β-LG) was expressed in Saccharomyces cerevisiae carrying bovine pre-β-LG cDNA and secreted into its growth medium. The expression plasmid was constructed by inserting the whole coding region of the cDNA encoding pre-β-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, β-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 β-LG secrected in the growth medium had the same mobility as that of authentic bovine β-LG. The N-terminal sequence was also identical with that of authentic mature bovine β-LG. |
| |
| Keywords: | |
|
|
