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Specific promoter ZFP6 of tobacco glandular hair and application of specific promoter

A specificity, promoter technology, applied in the field of bioengineering, can solve the problems of normal plant growth obstruction, metabolic balance disorder, etc., and achieve the effect of increasing transgene, improving aroma quality, and important theoretical significance

Inactive Publication Date: 2014-08-06
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Constitutive promoters can efficiently and non-specifically promote the expression of foreign genes in most or all plant tissues, but a large number of heterologous proteins will disturb the original metabolic balance in plants and hinder the normal growth of plants

Method used

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  • Specific promoter ZFP6 of tobacco glandular hair and application of specific promoter
  • Specific promoter ZFP6 of tobacco glandular hair and application of specific promoter
  • Specific promoter ZFP6 of tobacco glandular hair and application of specific promoter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 ZFP6 gene promoter fusion GUS reporter gene vector construction and tobacco genetic transformation

[0020] (1) The leaves of Arabidopsis thaliana were taken, and the genomic DNA was extracted by CTAB method.

[0021] (2) Design specific primers according to the upstream sequence of the Arabidopsis ZFP6 gene transcription initiation site on Genbank, and add SalⅠ and NotⅠ restriction enzyme sites to the 5‵ ends of the primers respectively. The sequences are as follows:

[0022] Primer 1: 5‵-AGTCGACTGCAACGTAAAAGGTCCTGA-3‵;

[0023] Primer 2: 5‵-AAGCGGCCGCCAAAGAGAGAGAGAGAGACAGATTT-3‵.

[0024] (3) PCR amplification was performed using the extracted DNA as a template.

[0025] high fidelity enzyme HS DNA Polymerase (TaKaRa Code: DR010A) system:

[0026] 5×PrimeSTAR Buffer (Mg 2+ plus) 10 μl,

[0027] dNTP Mixture (2.5mM each) 4μl,

[0028] Primer 1 (10 μM) 1 μl,

[0029] Primer 2 (10 μM) 1 μl,

[0030] Template DNA 1 μl,

[0031] HS DNA Polymerase 0.5...

Embodiment 2

[0049] The character observation of embodiment 2 transgenic tobacco

[0050] The epidermal hairs of tobacco are divided into two categories according to the presence or absence of secretory glands. One is the glandular hairs that can produce secretions, and the other is the protective hairs that do not have the ability to secrete. The glandular hairs are divided into long-stalked glandular hairs and short-stalked glands according to their shape. hair.

[0051] The transgenic tobacco obtained in Example 1 was subjected to histochemical staining (GUS staining) analysis, and the results were as follows: figure 1 As shown, after the 60-day-old tobacco leaves were stained with GUS, the promoters of the present invention (A, B) were specifically expressed in the glandular hairs of the tobacco leaves (the glandular hairs appeared blue), while the control (C, D ) has no color reaction, indicating that the promoter of the present invention is active in tobacco glandular trichomes and ...

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Abstract

The invention discloses a specific promoter ZFP6 of tobacco glandular hair and application of the specific promoter. The specific promoter or a complementary chain thereof has the nucleotide sequence as shown in SEQ ID NO.3. The invention further provides an application of the specific promoter in regulating the target gene expression in the tobacco glandular hair. The specific expression of the ZFP6 gene promoter in the glandular hair of tobacco leaves overcomes the wastes caused by the non-specific, continuing and efficient expression of exogenous genes promoted by constitutive promoters in a receiver plant, and enhances the transgenic effect; the specific promoter can be used for expressing the target gene in the tobacco glandular hair, and facilitates researching the regulation mechanism of the glandular hair forming and developing and the genetic regularity of the glandular hair density; on the basis of the intimate connection of the glandular hair and tobacco fragrance, the specific promoter is further conducive to researching the mechanism controlling the formation of the tobacco glandular hair and fragrance, and to improving the fragrance quality of the tobacco leaves, and has important theoretical meanings and potential application values.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, in particular to a tobacco glandular trichome-specific promoter ZFP6 and its application. Background technique [0002] Tobacco has become one of the most important economic crops in the global market in the 21st century. It is not only a source of income for tobacco farmers in more than 100 countries, but the entire tobacco industry involves different production departments and wholesale and retail links, and has become an important economic force in many countries. ; With the development of the tobacco industry, the local and local government taxes in many countries have been greatly improved. [0003] Tobacco is a high-efficiency crop. The planting area and output of tobacco in my country rank first in the world. Of all tobacco leaf types, flue-cured tobacco is the most widely grown. my country is the country that produces the most flue-cured tobacco in the world. The planting area an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/84A01H5/00
Inventor 甘银波颜安孟小芳安丽君周忠静赵永钦刘一华
Owner ZHEJIANG UNIV