Mulberry micromolecule heat shock protein gene Mn16.8-CI promoter and recombinant expression vector and application thereof

A technology of mn16.8-ci, heat shock protein, applied in the field of molecular biology, can solve the problem of less research on the mechanism of mulberry stress resistance

Inactive Publication Date: 2016-08-10
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few studies on the mechanism of mulberry stress resistance, especially how mulberry regulates the expression of downstream genes under high temperature stress.

Method used

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  • Mulberry micromolecule heat shock protein gene Mn16.8-CI promoter and recombinant expression vector and application thereof
  • Mulberry micromolecule heat shock protein gene Mn16.8-CI promoter and recombinant expression vector and application thereof
  • Mulberry micromolecule heat shock protein gene Mn16.8-CI promoter and recombinant expression vector and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1, Acquisition of Mn16.8-C Ⅰ promoter of Morus alba small molecule heat shock protein gene

[0020] 29 sHSPs were identified from the mulberry genome database. After analyzing the gene structure, transcriptional regulatory elements and tissue expression profiles of the 29 sHSPs, it was found that the promoter of the Mn16.8-C Ⅰ gene contained a complete heat shock element (HSE). Gene expression was upregulated under high temperature, low temperature, drought and high salt stress. In order to study the activity of the Mn16.8-C Ⅰ promoter, primers were designed from the upstream 1500bp promoter sequence of the Mn16.8-C Ⅰ gene, cloned and verified by sequencing. Using plant transgenic technology, insert it into the plant expression vector pBI121, the specific method is as follows:

[0021] According to the genome sequence of Chuanmulberry, the upstream 1500bp sequence of the Mn16.8-C Ⅰ gene was intercepted to design primers as primers for cloning the Mn16.8-C Ⅰ pro...

Embodiment 2

[0026] Example 2. Obtaining and Identification of Mn16.8-C Ⅰ Promoter Transgenic Tobacco

[0027] The constructed pBI121-Mn16.8-C Ⅰ expression vector was transformed into tobacco by leaf disk transformation method, and 5 transgenic tobacco plants were obtained after kan resistance screening, named T1-5 respectively. Genomic DNA was extracted from the leaves of 5 transgenic tobacco plants as a template, and the wild-type tobacco genome was used as a control, and PCR amplification was performed with the upstream primers of the Mn16.8-C Ⅰ promoter and the specific primers of the GUS gene. The sequences of the primers are as follows:

[0028] 5'-cgaggtacggtagaggttgg-3' (SEQ ID NO. 4);

[0029] PCR amplification conditions were pre-denaturation at 94°C for 4 minutes; denaturation at 94°C for 40 seconds, annealing at 60°C for 40 seconds, extension at 72°C for 2 minutes, and 30 cycles; final extension at 72°C for 10 minutes, and storage at 4°C; Agarose gel electrophoresis, the resu...

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Abstract

The invention discloses the Mn16.8-C I promoter of the mulberry small-molecule heat shock protein gene and its recombinant expression vector and application. Its nucleotide sequence is shown in SEQ ID NO.3, and the gene can be up-regulated downstream under high-temperature stress Gene expression, so the characteristics of the gene can be used to regulate the expression of the target gene through temperature regulation to regulate plant stress resistance or regulate disease resistance; at the same time, it provides important promoter elements for the study of plant stress resistance and disease resistance, and can also be used It is suitable for genetic engineering breeding of plant stress resistance, so it has wide application value.

Description

technical field [0001] The present invention belongs to the field of molecular biology, and specifically relates to the Mn16.8-C Ⅰ promoter of the mulberry small-molecule heat-shock protein gene, and also relates to a recombinant expression vector containing the Mn16.8-C Ⅰ promoter of the mulberry small-molecule heat-shock protein gene and the corresponding Applications. Background technique [0002] Plants growing in the natural environment are inevitably harmed by various environmental stresses. When subjected to adversity stress, plants initiate a stress response, and a series of changes occur in their morphological structure, physiology, biochemistry and molecular levels. During the long-term evolution process, plants have developed and formed a complex set of mechanisms to adapt to changes in the surrounding environment. [0003] Plants can rapidly synthesize a class of proteins—heat shock proteins (HSPs)—under high temperature stress. As molecular chaperones, HSPs c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/82
CPCC07K14/415C12N15/8273C12N15/8279
Inventor 何宁佳卢承琼曾其伟
Owner SOUTHWEST UNIV
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