Application of rice transcription factor Os05g39950 gene

A technology of n-linker-os05g39950, rice transcription factor, applied in the field of genetic engineering, can solve problems such as unseen OVATE protein regulation mechanism

Active Publication Date: 2012-11-21
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Application of rice transcription factor Os05g39950 gene
  • Application of rice transcription factor Os05g39950 gene
  • Application of rice transcription factor Os05g39950 gene

Examples

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Embodiment 1

[0025] Example 1 Obtaining of Rice Transcription Factor Os05g39950 Gene and Construction of Plant Expression Vector

[0026] Log in to the http: / / rice.plantbiology.msu.edu / analyses_search_locus.shtml website, find the rice transcription factor Os05g39950 gene, and design PCR amplification primers based on its sequence:

[0027] Os05g39950-F: 5'-CAAAAAAGCAGGCTTCATGTTGTCCAGCGAACCAGG-3'

[0028] Os05g39950-R: 5'-CAAGAAAGCTGGGTCTGGCATGACGCCACAGG-3'

[0029] The leaves of rice 'Nipponbare' were used as materials, total RNA was extracted by TRIzol method, and cDNA was obtained by reverse transcription. The reaction steps were as follows: (1) Add the following substances sequentially on ice to a PCR reaction tube without nuclease: 6 μl of total RNA ( 0.1ng-5μg), Oligo(dT)18 primer 1μl, nuclease-free ddH 2 O 5 μl, placed in a PCR instrument at 65°C for 5 minutes; (2) Then add the following substances: 4 μl of 5× reaction buffer, 1 μl of RNase inhibitor, 2 μl of 10 mM dNTP, 1 μl of M...

Embodiment 2

[0032] The acquisition of embodiment 2 transgenic rice plants

[0033] Take the mature seeds of rice 'kitaake', shell them manually or mechanically, select the plump, smooth and spot-free seeds and inoculate them on the induction medium for induction culture after being sterilized. Rice callus with good appearance and good growth was selected as the recipient material, and ubi:Os05g39950-VP64 was transferred into the rice callus by the Agrobacterium-mediated method. The AAM transformation liquid of Agrobacterium was transformed, and the callus soaked in the transformation liquid was placed on the co-culture medium for co-cultivation, cultured in the dark at 25°C for 3 days, then placed on the screening medium for about 30 days, and subcultured every 10 days. Then, the screened out resistant calli were transferred to the differentiation medium for differentiation for about 20 days, and subcultured every 10 days. The resistant calli that differentiated into green seedlings were...

Embodiment 3

[0038] Example 3 Identification of Os05g39950-VP64 transgenic rice

[0039] In order to detect whether the CDS sequence of the Os05g39950 gene is integrated into the rice genomic DNA, the total DNA of wild-type and transgenic rice leaves was extracted by CTAB method, and the detection primer sequence was designed. The upstream primer: 5'-ATGGACGCGCTGGACGATTT-3', the downstream primer: 5 '-TCACTTGTCATCGTCGTCCTTGTAG-3', for PCR, the amplification system is: reaction buffer 5μl, dNTP 1μl, ddH 2 O 2.2 μl, Taq DNA polymerase 0.2 μl, upstream and downstream primers 0.3 μl, DNA template 1 μl. The reaction conditions were hot start at 96°C for 5 minutes; Extend for 10 minutes, and finally finish the reaction at 25°C. The transgenic plants were identified, and the amplified products were subjected to agarose gel electrophoresis. The electrophoresis results showed that the target bands were amplified in the transgenic rice, but the target band was not amplified in the wild-type rice ( ...

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Abstract

The invention relates to application of rice transcription factor Os05g39950 gene. According to the application, a transcription factor activation sequence motif VP64 (four transcription factor activation sequence motifs VP16) and the rice transcription factor Os05g39950 gene are fused and built to obtain a forming transcription factor, and the forming transcription factor is converted into crops such as rice, so the rice grain property is improved, for example, the rice grain is obviously shortened, widened and thickened, and the grain weight is reduced. The important theoretical value is realized on the detail clarification of seed development mechanism regulation and control, and the rice grain form can be improved through a transgenosis measure, so the important significance is also realized in the production practice.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to the application of rice transcription factor Os05g39950 gene. Background technique [0002] Rice (Oryza sativa L.) is one of the most important food crops that humans depend on for survival, and it is also an important model species for functional gene research. The related genetics and molecular biology research has always attracted much attention. Among them, transcription Horizontal regulation is an important way to regulate gene expression. In the plant kingdom, plants that can form seeds account for more than two-thirds of the total number of plants. As important reproductive organs, seeds also provide food sources for people. Rice is an important representative of them. The seeds come from fertilized ovules . From the perspective of molecular biology, the development and germination of seeds is an orderly and selective process of gene expression. Transcript...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00
Inventor 刘军于慧边鸣镝李宏宇赵涛刘斌林辰涛
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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