.alpha.-lactalbumin gene constructs
AssigneePPL Therapeutics (Scotland) Limited
InventorsJulian David Cooper, Angelika Elisabeth Schnieke
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Abstract
The present invention utilizes genetic engineering techniques to prepare non-human transgenic mammals that express human .alpha.-lactalbumin in their milk at a concentration of 2 mg/ml or greater. The invention also includes methods of preparing human .alpha.-lactalbumin in, for example, mice and cows. Also taught are methods for preparing human .alpha.-lactalbumin in which from one to four of its natural phenylalanine residues have been substituted by another amino acid.
Claims5
A method of producing non-human animal milk containing at least 2 mg/ml of an .alpha.-lactalbumin selected from the group consisting of:
- (i) human .alpha.-lactalbumin;
- and
- (ii) modified human .alpha.-lactalbumin having from one to four of the natural phenylalanine residues substituted by other amino acid residues;
- said method comprising:
- (1) producing milk in the mammary gland of an adult transgenic, non-human placental mammal whose somatic and germ cells include a genetic construct comprising, in the 5' to 3' direction and operatively linked:
- (a) at least 1.8 kb of 5'-flanking sequence from the human .alpha.-lactalbumin gene including the .alpha.-lactalbumin promoter;
- (b) a DNA sequence encoding
- (1) a secretion signal;
- and
- (2) an .alpha.-lactalbumin selected from the group consisting of:
- (i) human .alpha.-lactalbumin;
- and
- (ii) modified human .alpha.-lactalbumin having from one to four of the natural phenylalanine residues substituted by other amino acid residues;
- (c) at least about 3 kb of 3'-flanking sequence from the human .alpha.-lactalbumin gene;
- wherein said construct is expressed in the mammary gland of said mammal and .alpha.-lactalbumin is produced in the milk at a level of at least 2 mg/ml;
- and
- (2) collecting the milk produced in step (1), wherein said milk contains at least 2 mg/ml of said human .alpha.-lactalbumin or said modified human .alpha.-lactalbumin.
A transgenic non-human mammal whose somatic and germ cells contain a transgene construct, said transgene construct comprising, in the 5' to 3' direction and operatively linked:
- (a) at least about 1.8 kb of 5'-flanking sequence from the human .alpha.-lactalbumin promoter:
- (b) a DNA sequence encoding
- (1) a signal sequence;
- and
- (2) a .alpha.-lactalbumin selected from the group consisting of:
- (i) human .alpha.-lactalbumin;
- and
- (ii) a modified human .alpha.-lactalbumin having from one to four of the natural phenylalanine residues substituted by other amino acid residues;
- (c) at least about 3 kb of 3'-flanking sequence from the human .alpha.-lactalbumin gene;
- wherein said transgene construct is integrated into the genome of said mammal in such a way that said DNA sequence is expressed in the mammary gland of said mammal to produce .alpha.-lactalbumin in the milk of said mammal at a level of at least 2 mg/ml.
Dependencies are inferred from the claim text.