Mammary gland specificity expression vector capable of efficiently expressing para-insulin growth factor IGF-1

A growth factor and expression vector technology, applied in the field of mammary gland-specific expression vector, can solve the problems of laborious, time-consuming breeding technology, insufficient market share, etc.

Inactive Publication Date: 2011-05-04
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the yield per head of sheep is relatively low, and the market share of goat milk is less than 5%, so increasing the production of goat milk has become an urgent task in front of us.
Conventional breeding techniques are time-consuming and labor-intensive; however, transgenic technology can quickly change a certain trait of a species

Method used

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  • Mammary gland specificity expression vector capable of efficiently expressing para-insulin growth factor IGF-1
  • Mammary gland specificity expression vector capable of efficiently expressing para-insulin growth factor IGF-1
  • Mammary gland specificity expression vector capable of efficiently expressing para-insulin growth factor IGF-1

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Experimental program
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Embodiment Construction

[0035] The methods used in the following examples are conventional methods unless otherwise specified.

[0036] The specific materials and reagents involved are as follows:

[0037] Sheep liver tissue comes from Saanen dairy goat with no history of infection (purchased from Nanjing Lishui Goat Farm), and the tissue is frozen in liquid nitrogen, and the skeleton vector pBC1 (Company: Invitrogen; Catalog Number: K27001; Product: pBC1Milk Expression Vector Kit ); vector pCDNA3.1 (Company: Invitrogen; English name: PCDNA3.1 / ZEO(+); Catalog Number: V86020); human breast cancer cell Bcap-37 (Nanjing Kaiji Biological Company; catalog number KG035; name: human breast Cancer cell Bcap-37); vector pBluescriptIISK (+) and E.coli JM109 (purchased from Dalian Baobio, Takara).

[0038] Reagents TRizol, MLV-Reverse Transcriptase, rTaq Enzyme, pMD19-T Vector, Agarose Gel DNAPurification Kit, In-Fusion TMAdvantage PCR Cloning Kit, TaKaRa DNA Ligation Kit endonucleases Xho I, Not I, Kpn I, Cl...

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Abstract

The invention discloses a mammary gland specificity expression vector capable of efficiently expressing para-insulin growth factor IGF-1, belonging to the technical field of biological engineering. The cDNA sequence of the IGF-1 is disclosed as SEQ ID NO.1. The mammary gland specificity expression vector pIN is transfected to the human mammary gland cell Bcap-37 in vitro by using a liposome method, and albumin and cell protein on the cell are extracted; and the contents of the IGF-1 in the albumin and cell protein on the cell are detected by using a western blot method and a radio-immunity method. The result indicates that the expression amount of the IGF-1 in the group transfected by the mammary gland specificity expression vector pIN is obviously increased as compared with the control group which is not transfected by the mammary gland specificity expression vector pIN. Thus, the invention indicates that the pIN vector can successfully express the IGF-1 on the mammary gland cell, thereby laying the solid foundation for the production of the IGF-1-modified milk goat in the next step.

Description

1. Technical field [0001] The invention relates to a mammary gland-specific expression vector for highly expressing insulin-like growth factor, which belongs to the field of biotechnology, and in particular relates to insulin-like growth factor IGF-1 for promoting lactation and increasing milk production. 2. Background technology [0002] Goat milk is rich in nutrition and is the most "close to perfect food"; it is called "white blood" and is the most ideal natural food. However, the yield per head of sheep is relatively low, and the market share of goat milk is less than 5%, so increasing the yield of goat milk has become an urgent task before us. Conventional breeding technology is time-consuming and labor-intensive; while transgenic technology can just change a certain trait of a species quickly. The main factors affecting mammary gland development in vivo are growth hormone, prolactin, progesterone, estrogen and insulin-like growth factor (IGF-1). Among them, IGF-1 pla...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N15/12C12N15/64
Inventor 杨倩林建庾庆华张强
Owner NANJING AGRICULTURAL UNIVERSITY
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