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Application of cds sequence of rice transcription factor os05g10670.1 gene

A rice transcription factor and transcription factor technology, applied in the field of genetic engineering, can solve the problems of weakening of hybrid breeding advantages

Active Publication Date: 2018-05-01
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current research on increasing rice production is more dependent on limited rice germplasm resources, the advantages of traditional hybrid breeding are gradually weakening, and rice transgenic technology may explore the potential of further increasing rice production

Method used

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  • Application of cds sequence of rice transcription factor os05g10670.1 gene
  • Application of cds sequence of rice transcription factor os05g10670.1 gene
  • Application of cds sequence of rice transcription factor os05g10670.1 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Isolation of Os05g10670.1 Gene CDS Sequence and Plant Expression Vector Construction

[0034] 1. Acquisition of CDS sequence of Os05g10670.1 gene

[0035]Find the Os05g10670.1 gene in the plant transcription factor database (http: / / rice.plantbiology.msu.edu / analyses_search_locus.shtml), design PCR amplification primers according to its CDS sequence, and design PCR amplification primers according to its sequence (F: 5′-CAAAAAAGCAGGCTTCATGGTTCGGAAGCGTCG-3′ and reverse primer R: 5′-CAAGAAAGCTGGGTCGTTGATGAGGTCGGATACCC-3′). The CDS sequence of the Os05g10670.1 gene was obtained by PCR using the total cDNA of wild-type Nipponbare rice as a template, and its nucleotide sequence is shown in SEQ ID No.1.

[0036] 2. Construction of plant expression vectors

[0037] The CDS sequence of the rice transcription factor Os05g10670.1 gene was constructed downstream of the 4 transcription factor activation motif VP16 coding gene (SEQ ID No.4) through the Gateway system.

[...

Embodiment 2

[0047] The acquisition of embodiment 2 transgenic rice plants

[0048] 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: VP64-Os05g10670.1 was transferred into the rice callus by Agrobacterium-mediated method, with 100 μM acetosyringone and O.D. 0.7 AAM culture solution of Agrobacterium. For transformation, the callus soaked in the transformation solution was placed on a co-culture medium for co-cultivation, cultured in the dark at 25°C for 3 days, then placed on a 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 differentia...

Embodiment 3

[0054] Identification of embodiment 3 transgenic positive strains

[0055] In order to detect the overexpression of the ubi:VP64-Os05g10670.1 gene in the VP64-Os05g10670.1 transgenic rice line obtained in Example 2 in the T2 transgenic rice (V1430H-04, V1430H-14), this example was Primers were designed on the vector (forward primer: 5'-GTGGGGACAAGTTTGTACAAAAAGCAGGCTTC-3' and reverse primer: 5'-GTGGGGACCACTTTGTACAAGAAAGCTGGGTC-3') for PCR detection, and obvious specific bands were obtained. Depend on image 3 It can be seen that the transgenic rice lines V1430H-04 and V1430H-14 obtained in Example 2 contain the VP64-Os05g10670.1 fusion gene. The above primers are designed at the junction of the vector and the target gene, image 3 It shows that the size of the amplified fragment is basically consistent with the target gene (1395bp)

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Abstract

The present invention relates to the application of the CDS sequence of the rice transcription factor Os05g10670.1 gene, which uses the fusion of the transcription factor activation motif VP64 and the rice transcription factor Os05g10670.1 gene CDS sequence to construct a constitutive transcription factor, and transforms it into crops such as rice, Thereby improving rice grain traits, such as increasing rice grain length. It has important theoretical value for elucidating the mechanism of regulating seed development in detail, and can improve the grain shape of rice through transgenic means, so it is also of great significance in production practice.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to the application of the rice transcription factor Os05g10670.1 gene CDS sequence. Background technique [0002] Rice (Oryza sativa L.) is one of the three most important food crops in my country and the world, the staple food of more than half of the world's population, and an important model plant for functional gene research. Related genetics and molecular biology studies have been paid much attention by researchers, and the regulation of transcription level is an important way of gene expression regulation. The current research on increasing rice yield is more dependent on limited rice germplasm resources, the advantages of traditional hybrid breeding are gradually weakening, and rice transgenic technology may explore the potential of further increasing rice yield. [0003] In the plant kingdom, plants that can form seeds account for more than two-thirds of the total number ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/82C12N1/21A01H5/10A01H6/46C12N15/11
Inventor 刘斌李宏宇赵涛刘军张春雨林辰涛
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI