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.

Inactive Publication Date: 2011-03-16
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

Compared with cow milk, the size of fat particles in goat milk is only 1/3 of that in milk. The size of fat particles in goat milk is uniform, the content of unsaturated fatty acids is high, and it is in a good emulsified state. The content of goat milk milk protein is high, an...

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  • Mammary gland specificity expression vector efficiently expressing goat growth hormone (GH)
  • Mammary gland specificity expression vector efficiently expressing goat growth hormone (GH)
  • Mammary gland specificity expression vector efficiently expressing goat growth hormone (GH)

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Embodiment Construction

[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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Abstract

The invention discloses a mammary gland specificity expression vector efficiently expressing a goat growth hormone (GH) and belongs to the technical field of biological engineering. The cDNA sequence of the GH is shown as SEQ ID NO. 1. The growth hormone can promote the formulation of a mammary duct and accelerate the proliferation of mammary epithelial cells of a goat so as to increase milk yield. The mammary gland specificity expression vector pcGH constructed by the invention is introduced into a human breast cancer cell Bcap-37 cultured in vitro by a liposome method, and cell culture supernatant is collected after transfection; and the GH content in the cell culture supernatant is detected by an enzyme-linked immuno sorbent assay (ELISA) method. A result shows that the GH expression amount of the cells in a pcGH-transfected group is obviously higher than that of the cells in a blank group which is not transfected by the pcGH. And the result shows that the pcGH vector successfully expresses the GH on mammary cells, so that a solid foundation for breeding a gh-transfected genic milk goat in the next step is laid.

Description

1. Technical Field [0001] The invention relates to a mammary gland specific expression vector for efficiently expressing goat growth hormone GH, which belongs to the field of biotechnology, and specifically relates to growth hormone GH that promotes the development of mammary ducts and increases milk production. 2. Background technology [0002] Goat milk is an important part of our country's dairy industry. Goat milk has small fat particles and is rich in protein, minerals, vitamins and biologically active substances. Compared with cow’s milk, the size of fat particles in goat’s milk is only 1 / 3 of that in milk. The fat particles in goat’s milk are even in size, have a high content of unsaturated fatty acids, and are in a good state of emulsification. Goat’s milk has high protein content and has a similar protein structure. Breast milk is the same, and the protein clot is fine and soft, which is more conducive to human absorption. Although my country is a big country of dairy goa...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/18C12N15/12C12N15/64
Inventor 杨倩张强庾庆华林建
Owner NANJING AGRICULTURAL UNIVERSITY
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