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Mammary gland specificity expression vector and construction method thereof

An expression vector and specific technology, applied in the field of mammary gland-specific expression vector and its construction, can solve the problem of low endogenous expression level

Active Publication Date: 2010-09-29
INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, the research results are only limited to the expression of the milk protein BAC locus, and people are more interested in how to use the regulatory sequence in the milk protein BAC locus to guide the expression of foreign genes, especially the expression of some important medical genes. Only in this way can the milk protein loci be truly utilized, making the animal mammary gland bioreactor a versatile, convenient and effective expression system
To achieve this goal, the key to the problem is how to rationally assemble the coding sequence of the target gene to be expressed and the regulatory sequence in the milk protein locus, and the research in this area is still in its infancy

Method used

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  • Mammary gland specificity expression vector and construction method thereof
  • Mammary gland specificity expression vector and construction method thereof
  • Mammary gland specificity expression vector and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1. Construction of Human Serum Albumin Mammary Gland-Specific Expression Vector Using Sheep β-Casein Locus

[0041] 1. Homologous arm cloning of goat β-casein-human serum albumin and construction of capture vector for three consecutive times

[0042] Three pairs (6 in total) of homology arms with a length of 400bp-600bp, in which the first pair of homology arms are respectively located at the 3' end of the sheep β-casein locus and near the stop codon of the sheep β-casein genome sequence , used to capture the 3' regulatory region of the sheep β-casein locus; the second pair of homology arms are located near the start codon and stop codon of the human serum albumin gene genome sequence, used to capture the complete human The genome sequence of the serum albumin gene; the third pair of homology arms are located at the 5' end of the sheep β-casein locus and near the start codon of the sheep β-casein genome sequence, used to capture the sheep β-casein The 5' regula...

Embodiment 2

[0118] Example 2. Construction of Human Serum Albumin Mammary Gland-Specific Expression Vector Using the Bovine β-Casein Locus

[0119] 1. Homology arm cloning of bovine β-casein-human serum albumin and construction of capture vector for three consecutive times

[0120] Three pairs (6 in total) of homology arms with a length of 400bp-600bp, wherein the first pair of homology arms are respectively located at the 3' end of the bovine β-casein locus and near the stop codon of the bovine β-casein genome sequence, Used to capture the 3' regulatory region of the bovine β-casein locus; the second pair of homology arms are located near the start codon and stop codon of the human serum albumin gene genome sequence, used to capture intact human serum Genomic sequence of the albumin gene; the third pair of homology arms are located at the 5' end of the bovine β-casein locus and near the start codon of the bovine β-casein genomic sequence, for capturing the bovine β-casein gene The 5' re...

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Abstract

The invention discloses a mammary gland-specific expression vector and a construction method thereof. The construction method of the mammary gland-specific expression vector provided by the method comprises the following steps: (1) a DNA fragments that are M0 are constructed; the M0 is T1 to T6 that are homologous arms in sequence from upstream position to downstream position; (2) the M0 is inserted into the vector, and a continuous three-time gene capturing vector is obtained; (3) T5 and T6 capture a 3<1> regulatory region of lactoprotein loca through homologous recombinations for three times; T3 and T4 capture an integral exogenous proteinic genome sequence; T1 and T2 capture a 5<1> regulatory region of the lactoprotein loca. The method gets a lactoprotein-objective protein heterozygousloca, an upstream expression regulation sequence and a downstream expression regulation sequence of the lactoprotein are contained in the heterozygous loca, and the exogenous proteinic genome sequence is contained in the middle part. The mammary gland-specific expression vector and the construction method thereof inject new energies into the individual expression system of animal mammary glands, and greatly promote the further application of a mammary gland bioreactor expression system.

Description

technical field [0001] The invention relates to a mammary gland-specific expression vector and a construction method thereof. Background technique [0002] At present, the commonly used protein expression systems mainly include bacteria, yeast, mammalian cells, animal bioreactors, etc. The above-mentioned protein expression systems have their own advantages, disadvantages and scope of application. Proteins with different characteristics need to choose corresponding expression systems. [0003] Animal mammary gland bioreactor (mammary gland bioreactor) is an important branch of animal bioreactors, also known as animal individual mammary gland expression system, which belongs to the category of transgenic animals, and its core content is to use various transgenic techniques to specific mammary gland tissue The exogenous gene driven by sex promoter is highly expressed in animal mammary gland tissue, and the target product is produced in milk. How to efficiently, stably, and sp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/85C12N15/14C12N15/11C12N5/10C12N1/15C12N1/19C12N1/21A01K67/027C12N15/113C12N15/66
Inventor 陈红星施庚寿吴晓洁周颜荣黄培堂
Owner INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE