Mammary gland specificity expression vector efficiently expressing goat growth hormone (GH)
A growth hormone and expression vector technology, which can be used in the introduction of foreign genetic material using vectors, biochemical equipment and methods, applications, etc., and can solve problems such as difficulty in meeting market demand and low milk yield.
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[0036] The present invention first amplifies the target gene goat gh and goat β-lactoglobulin 5 and 3 end genes, and then uses the mammary gland specific expression vector pIRES 2 -EGFP is the basis to construct the breast-specific expression vector pcGH, and finally the human breast cancer cell Bcap-37 is transfected by liposome transfection method, and the protein is extracted by ELISA method to detect the expression effect after transfection.
[0037] The materials and reagents involved are as follows:
[0038] Sheep pituitary gland, from Saanen dairy goat without a history of infection, preserved in liquid nitrogen (purchased from Nanjing Lishui goat breeding base), pIRES 2 -EGFP vector (purchased from Clontech); human breast cancer cell Bcap-37 (Nanjing KGI Biotech Company; item number KG035; name: human breast cancer cell Bcap-37), vector pMD19-T, strain JM109 (purchased from Dalian Biotech , Takara).
[0039] Reagents Trizol, MLV-reverse transcriptase were purchased from Prome...
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