Method for constructing idiosyncratic carrier of galactophore of transgenic animal in high expression level

A technique for transgenic animals and construction methods, which is applied in the field of mammary gland bioreactors for transgenic animals, can solve the problem that milk protein regulatory sequences are difficult to reach a high level, and achieve great application value, increase production, and improve economic benefits

Inactive Publication Date: 2006-12-06
YANGZHOU UNIV
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  • Abstract
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Problems solved by technology

[0003] At present, the main problem to be solved for animal mammary gland-specific expression is still the expression le

Method used

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Embodiment Construction

[0017] 1. Gene construction process:

[0018] 1. The cytomegalovirus enhancer sequence is derived from the pcDNA vector, and its fragment is taken. The gene fragment has the following sequence characteristics:

[0019] cgttacataa ccttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 60

[0020] gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 120

[0021] atgggtggag tattatacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 180

[0022] aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 240

[0023] catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 300

[0024] catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 360

[0025] atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 420

[0026] ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 480

[0027] acggtggggag gtctatataa gcagagctct caattata

[0028] 2. The cytomegalovirus enhancer sequence is constructed between the 4-10kb upstr...

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Abstract

This invention discloses a method for constructing animal mammary gland-specific vector with high expression level. The method comprises: (1) utilizing partial non-coding sequence (about 500bp) of cytomegalovirus as an enhancer; (2) inserting the enhancer between a lactoprotein regulatory sequence and a functional gene (Cdna or genome DNA) to construct a mammary gland-specific expression vector. The regulatory sequence is non-coding sequence of casein or lactoglobulin. The specific expression vector can increase the expression level and the expression efficiency of a transgenic animal mammary gland bioreactor.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and the technology is applied to prepare transgenic animal mammary gland bioreactors. Background technique [0002] Transgenic animal mammary gland bioreactor is to use animal mammary gland organs to produce biological drugs or biological products, which has high commercial application value, but the efficiency in the preparation of mammary gland bioreactor is low, and the expression rate and expression level in transgenic animal mammary gland are Random and indeterminate, most were expressed at low levels. At present, the general mammary gland-specific expression gene is composed of milk protein regulatory element and functional gene. Although this gene component has a regulatory effect on the initiation and specificity of functional gene expression, its expression level and expression rate are very low. Its overall gene structure is: 5'-terminal milk protein regulatory sequence→function...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/33C12N15/67
Inventor 成勇
Owner YANGZHOU UNIV
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