Methods and vectors for improving nucleic acid expression
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Abstract
The invention features methods of making transgenic animals, and transgenic animals made by such methods. The method includes introducing into a cell, a nucleic acid construct comprising a nucleic acid sequence encoding a heterologous polypeptide under the control of a mammary epithelial cell promoter, an insulator positioned 5′ from the promoter, an insulator positioned 3′ from the nucleic acid sequence encoding the polypeptide, and a prokaryotic sequence, wherein the sequence between the insulator 5′ from the promoter and the insulator 3′ from the nucleic acid encoding the polypeptide is substantially free of prokaryotic sequence; and allowing a transgenic mammal to develop from the cell, to thereby provide a transgenic mammal.
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Citations
32 Claims
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1-26. -26. (canceled)
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27. A method of making a transgenic non-human mammal, comprising:
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introducing into a cell, a nucleic acid construct comprising a nucleic acid sequence encoding a heterologous polypeptide under the control of a mammary epithelial cell promoter, an insulator positioned 5′
from the promoter, an insulator positioned 3′
from the nucleic acid sequence encoding the polypeptide, and a prokaryotic sequence, wherein the sequence between the insulator 5′
from the promoter and the insulator 3′
from the nucleic acid encoding the polypeptide is substantially free of prokaryotic sequence;
allowing a transgenic mammal to develop from the cell, to thereby provide a transgenic mammal;
wherein substantially free of prokaryotic sequence means that the nucleic acid sequence has 0.5 kb or less of prokaryotic sequence between the insulator positioned 5′
from the promoter and the insulator sequence positioned 3′
from the nucleic acid sequence encoding the polypeptide; and
,wherein the nucleic acid sequence encoding the heterologous polypeptide under the control of a mammary epithelial cell promoter is genetically engineered into the genome of a somatic cell which is later fused with a enucleated oocyte to form a nuclear transfer unit. - View Dependent Claims (28, 29, 30, 31, 32)
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Specification