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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 low milk yield and difficulty in meeting market demand.

Inactive Publication Date: 2012-06-13
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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, and the protein structure is similar to The same as breast milk, the protein clots are thin and soft, which is more conducive to human body absorption. Although my country is a big country for raising dairy goats, the milk production is low and it is difficult to meet the market demand

Method used

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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 gh of goat and the 5-end and 3-end genes of goat β-lactoglobulin, and then uses the mammary gland-specific expression vector pIRES 2 -EGFP was used as the basis to construct the mammary gland-specific expression vector pcGH, and finally, human breast cancer cell Bcap-37 was transfected by liposome transfection method, and the protein was extracted by ELISA method to detect the expression effect after transfection.

[0037] The specific materials and reagents involved are as follows:

[0038] Sheep pituitary gland, from a Saanen dairy goat with no history of infection, was preserved in liquid nitrogen (purchased from Nanjing Lishui Breeding Goat Breeding Base), pIRES 2 -EGFP carrier (purchased from Clontech); human breast cancer cell Bcap-37 (Nanjing Kaiji Biological Company; product number KG035; name: human breast cancer cell Bcap-37), carrier pMD19-T, JM109 strain (purchased from Dalian Baobio , Takara).

[0039]...

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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 a growth hormone GH for promoting the development of mammary gland ducts and increasing milk production. 2. Background technology [0002] Goat milk is an important part of my country's dairy industry. Goat milk fat particles are small in size and rich in protein, minerals, vitamins and biologically active substances. 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, and the protein structure is similar to The same as breast milk, the protein clots are thin and soft, which is more conducive to human absorption. Although my country is a big ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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