Secreted Proteins and Nucleic Acids Encoding Them
AssigneeMillennium BioTherapeutics, Inc.
InventorsSean A. McCarthy, Steven Bossone
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Abstract
The invention involves a method of identifying nucleic acid sequences encoding signal peptide-containing proteins. The method features chimeric constructs containing a KRE9 gene that lacks a signal sequence. Yeast containing chimeric KRE9 plasmid constructs that encode signal sequences are selected based on their ability to grow on media in which sucrose is the sole carbon source.
Claims17
A method comprising:
- (a) obtaining a nucleic acid molecule comprising a chimeric gene, said chimeric gene comprising a first portion and a second portion, the first portion encoding a KRE9 lacking a functional signal sequence and the second portion being a heterologous nucleic acid sequence;
- (b) transforming a yeast cell lacking a functional KRE9 gene with said nucleic acid molecule;
- and
- (c) determining whether said transformed yeast cell grows when supplied with a medium that permits growth of a yeast cell expressing KRE9 having a functional signal sequence, but does not permit growth of a yeast cell that does not express KRE9 having a functional signal sequence, wherein growth on said medium indicates that said heterologous nucleic acid sequence present in said yeast cell encodes a signal sequence.
A yeast cell transformed with a nucleic acid molecule comprising a chimeric gene, said chimeric gene comprising a first portion and a second portion, the first portion encoding a KRE9 lacking a functional signal sequence and the second portion being a heterologous nucleic acid sequence.
A method comprising:
- (a) obtaining a nucleic acid molecule comprising a chimeric gene, said chimeric gene comprising a first portion and a second portion, the first portion encoding a KRE9 lacking a functional signal sequence and the second portion being a heterologous nucleic acid sequence;
- (b) transforming a yeast cell lacking a functional KRE9 gene with said nucleic acid molecule;
- and
- (c) determining whether said transformed yeast cell grows when supplied with a medium that does not permit growth of a yeast cell expressing KRE9 having a functional signal sequence, but does permit growth of a yeast cell that does not express KRE9 having a functional signal sequence, wherein lack of growth on said medium indicates that said heterologous nucleic acid sequence present in said yeast cell encodes a signal sequence.
The expression vector pBOSS-1.
Dependencies are inferred from the claim text.