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Construction of La-tPA vector and expression in the mammary gland of transgenic mice.

Yifan Lu, Jixian Deng, Xuan Cheng, Jiang Feng Zhou, Peitang Huang

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Abstract

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.

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What this paper is about

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.

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Available abstract

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.

Key concepts: Microinjection, Transgene, Molecular biology, Mammary gland, Southern blot, Genetically modified mouse, Northern blot, Plasminogen activator

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Construction of La-tPA vector and expression in the mammary gland of transgenic mice. — Research Paper | ScholarLens