Construction of yeast secretion vectors designed for production of mature proteins using the signal sequence of yeast invertase

✍ Scribed by Masafumi Nishizawa; Fumiko Ozawa; Fumio Hishinuma

• Book ID: 104655037
• Publisher: Springer
• Year: 1989
• Tongue: English
• Weight: 990 KB
• Volume: 32
• Category: Article
• ISSN: 1432-0614
• DOI: 10.1007/bf00184982

✦ Synopsis

Two kinds of yeast secretion vectors were constructed by site-directed mutagenesis of the invertase signal sequence and ligation of synthetic oligonucleotides coding appropriate signals. Each has a cloning site for a foreign gene preceded by a sequence encoding either the signal peptide cleavage site or a Lys-Arg sequence which is a cleavage site for the product of the KEX2 gene. Both vectors were able to direct the expression and secretion of mouse amylase. One of them has a Sall site within the signal sequence, and an attempt to clone sequences enhancing secretion of amylase with this vector is reported.

tase, which could be unfavourable for proper functioning of the product. To prevent the synthesis of the extension, vectors with a cloning site either at the signal cleavage site or at a position following a Lys-Arg sequence, which is a target site of the KEX2 gene product (KEX2p; Julius et al. 1984a), were constructed by site-directed mutagenesis and ligation of appropriate synthetic oligonucleotides. These vectors were able to direct the production and secretion of a foreign protein. The vectors proved useful for the study of factors affecting secretion efficiency.