Construction of La-tPA vector and expression in the mammary gland of transgenic mice.
Expression vectors of long acting human tissue plasminogen activator (La-tPa) in the mammary gland was constructed using the promoters of sheep beta-lactoglobulin gene (BLG, 5 kb) containing the first and the second introns obtained by PCR. Transgenic mice were established by microinjection. 540 fertilized eggs were injected and 6 transgenic mice were screened out from the offspring by PCR and Southern Blot analysis. The foreign gene integrating rate was 32%. The expression of La-tPA in mammary glands of transgenic mice was confirmed by milk assay and Northern blot analysis. Expression level of La-tPA reached 6 ug/ml in milk of transgenic mice.
Lu
Y
Y
Deng
J
J
Cheng
X
X
Zhou
J
J
Huang
P
P
D000818:Animals; D015152:Blotting, Northern; D005260:Female; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D007438:Introns; D007782:Lactoglobulins / Q000235:genetics; D008321:Mammary Glands, Animal / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D008892:Milk / Q000378:metabolism; D010960:Plasminogen Activators / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012756:Sheep
D007782:Lactoglobulins; D010960:Plasminogen Activators