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AssigneeNational University of Singapore
InventorsJeak Ling Ding, Nguan Soon Tan, Bow Ho, Toong Jin Lam
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A universal secretory signal originally derived from a piscine vitellogenin (Vtg) gene is inserted into various expression vectors for driving the secretion of the recombinant protein into the culture medium. This enhances the detection, quantification and downstream scaled-up purification of a recombinant protein of interest. The secretory signal system is very versatile, being conveniently and widely applicable to an array of heterologous host cells such as bacteria, yeast, insect, piscine, and mammalian cell lines (e.g., COS, CHO, NIH/3T3). The said secretory system is also applicable as a reporter vector for secretion of reporter proteins/enzymes, thus, enabling the detection of the reporter proteins (e.g., CAT, GFP) in the culture medium.
An isolated nucleic acid comprising a nucleotide sequence encoding a secretory signal sequence comprising the amino acid sequence SEQ ID NO:10, or variants of said amino acid sequence that comprise conservative replacements thereof that retain the biological activities of directing secretion of a fusion protein from a cell and cleavage of the secretory signal sequence from the fusion protein, wherein the variations in said variants
An isolated nucleic acid comprising a nucleotide sequence encoding a fusion protein comprising a secretory signal sequence and a desired heterologous protein, wherein said secretory signal sequence, comprises the amino acid sequence SEQ ID NO:10, or variants of said amino acid sequence that comprise conservative replacements thereof that retain the biological activities of directing secretion of a fusion protein from a cell and cleavage of the secretory signal sequence from the fusion protein, wherein the variations in said variants
A fusion protein comprising
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