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Plant bidirectional promoter BIGDB2

A promoter and nucleotide sequence technology, applied in plant products, botanical equipment and methods, angiosperms/flowering plants, etc., can solve problems such as insertion

Active Publication Date: 2012-05-16
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0003] People have a relatively deep understanding of the phenomenon of gene silencing caused by transgenes. There are many factors that cause transgene silencing, the most important of which is the generation of repetitive sequences, especially when multiple genes are introduced, more artificially caused multiple 35S The case of promoter insertion, for which there is no effective solution so far

Method used

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  • Plant bidirectional promoter BIGDB2
  • Plant bidirectional promoter BIGDB2
  • Plant bidirectional promoter BIGDB2

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Experimental program
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Embodiment 1

[0029] Embodiment 1, the acquisition of promoter and its detection

[0030] 1. Acquisition of the promoter

[0031] According to the results of bioinformatics analysis, there are some potential bidirectional promoter sequences in the Arabidopsis genome. Using Arabidopsis genomic DNA as a template, the forward primer (BIGDB2-F: 5′-GAGCTCTAGA GCGGC CGC TCTAAACCCTAATTATCTCTCTTTCTCGTC-3′) and reverse primer (BIGDB2-R: 5′-ACGC GTC GAC CCCGGGCTTCTTCTTCGTCTACGAGAGTTCTTTG-3′) amplified, the amplified product was separated by agarose electrophoresis, recovered and connected to the pEASY-T1 (Quanshijin Company) cloning vector, transformed into Escherichia coli XL1-BLUE (Agilent Technologies-Stratagene Products), and passed After sequencing, it was confirmed that the cloning vector was correct, and the nucleotide sequence of the amplified product (named BIGDB2) was shown in sequence 1 in the sequence listing.

[0032] 2. Construction of recombinant expression vector

[0033] 1. C...

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Abstract

The invention discloses a plant bidirectional promoter BIGDB2. The nucleotide sequence of the promoter is shown as a sequence 1 in a sequence table. Experiments prove that: the BIGDB2 can promote glucuronidase (GUS) expression after being inserted into the space between GUS and luciferase (LUC) in a vector pMOA34-G / L, and the BIGDB2 can also promote the GUS expression after being inserted into the space between the GUS and the LUC in the pMOA34-G / L reversely, so that the BIGDB2 has a bidirectional promoting function in arabidopsis thaliana.

Description

technical field [0001] The invention relates to the cloning and application of a plant bidirectional promoter BIGDB2. Background technique [0002] It is the ultimate goal of plant genetic engineering to obtain new traits and stably inherit them by introducing exogenous genes. The cauliflower virus CaMV 35S promoter has been widely used as a stable and efficient exogenous promoter. However, during the transgenic process, the appearance of repeated sequences will cause gene silencing, especially when multiple genes are introduced, and this phenomenon will be more serious, thus seriously affecting the acquisition of transgenic plants and the stability of offspring. When two or more genes are introduced, the expression amount, expression time and position of these genes are usually required to be identical, so as to ensure the normal function of the transferred genes. It is difficult to meet such requirements by using the 35S promoter. These are problems to be solved urgently...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00
Inventor 谢旗夏然
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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