Transgenically produced non-secreted proteins

A method for providing a non-secreted protein in the milk of a transgenic mammal, comprising

• obtaining milk from a transgenic mammal selected from the group consisting of sheep, mice, pigs, cows, rabbits and goats having introduced into its germline a nucleic acid construct comprising:
• (a) a mammary epithelial specific promoter sequence,
• (b) a milk protein specific signal sequence,
• (c) a sequence other than a signal sequence which encodes a sufficient portion of the amino terminal coding region from the amino terminal end of a milk protein to allow secretion in the milk of a transgenic mammal of a non-secreted protein;
• and
• (d) a sequence encoding a non-secreted protein,
• wherein elements (a), (b), (c), and (d) are operatively linked in the order recited and the heterologous non-secreted protein is expressed and secreted into the milk of the mammal, thereby providing a non-secreted protein in the milk of the mammal.

An isolated nucleic acid construct which comprises:

• (a) a mammary epithelial specific promoter;
• (b) a milk protein specific signal sequence which can direct the secretion of a protein;
• (c) a sequence other than a signal sequence which encodes a sufficient portion of the amino terminal coding region from the amino terminal end of a milk protein to allow secretion in the milk of a transgenic mammal of a non-secreted protein;
• and
• (d) a sequence which encodes a non-secreted protein,
• wherein elements (a), (b), (c), and (d) are operatively linked in the order recited.

A method of making and secreting a non-secreted protein comprising:

• introducing into a mammal selected from the group consisting of sheep, mice, pigs, cows, rabbits and goats a nucleic acid construct which comprises:
• (a) a mammary epithelial specific promoter;
• (b) a milk protein specific signal sequence which can direct the secretion of a protein;
• (c) a sequence other than a signal sequence which encodes about 10% of the amino terminal coding region from the amino terminal end of a milk protein to allow secretion in the milk of a transgenic mammal of a non-secreted protein;
• and
• (d) a sequence which encodes a non-secreted protein,
• wherein elements (a), (b), (c), and (d) are operatively linked in the order recited;
• and
• expressing the protein, thereby producing the non-secreted protein in the milk of the transgenic mammal.

A method for providing a non-secreted protein in the milk of a transgenic mammal, comprising

• obtaining milk from a transgenic mammal selected from the group consisting of sheep, mice, pigs, cows, rabbits and goats having introduced into its germline a nucleic acid construct comprising:
• (a) a mammary epithelial specific promoter sequence,
• (b) a milk protein specific signal sequence,
• (c) a sequence other than a signal sequence which encodes about 10% of the amino terminal coding region from the amino terminal end of a milk protein to allow secretion in the milk of a transgenic mammal of a non-secreted protein;
• and
• (d) a sequence encoding a non-secreted protein,
• wherein elements (a), (b), (c), and (d) are operatively linked in the order recited and the heterologous non-secreted protein is expressed and secreted into the milk of the mammal, thereby providing a non-secreted protein in the milk of the mammal.

An isolated nucleic acid construct which comprises:

• (a) a mammary epithelial specific promoter;
• (b) a milk protein specific signal sequence which can direct the secretion of a protein;
• (c) a sequence other than a signal sequence which encodes about 10% of the amino terminal coding region from the amino terminal end of a milk protein to allow secretion in the milk of a transgenic mammal of a non-secreted protein;
• and
• (d) a sequence which encodes a non-secreted protein,
• wherein elements (a), (b), (c), and (d) are operatively linked in the order recited.