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Preparation method of iku2 gene of peony purpura

A purple spot peony and gene technology, applied in the field of bioengineering, can solve problems that have not been reported, and achieve the effects of improving transformation efficiency, improving size, and increasing plant yield

Active Publication Date: 2021-05-11
DALIAN NATIONALITIES UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the functional research of the IKU2 gene has only been verified in model plants such as Arabidopsis thaliana, and the function of the IKU2 gene in woody oil crops, especially the purple spot peony, has not been reported yet.

Method used

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  • Preparation method of iku2 gene of peony purpura
  • Preparation method of iku2 gene of peony purpura
  • Preparation method of iku2 gene of peony purpura

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Embodiment 1

[0046] 1. Acquisition of IKU2 gene cDNA from the seeds of Paeonia japonicus

[0047] The seeds of the four developmental periods (June 20, July 7, July 17 and July 27) were collected from the purple spot peony base of Dalian University for Nationalities, Jinshitan Campus, marked and immediately frozen in liquid nitrogen, and then the seeds were Grinding with liquid nitrogen, according to the recommended method of the column type plant total RNA extraction and purification kit of Shanghai Sangong Co., Ltd., the total RNA was extracted from the seeds of peony purpurea. Using the extracted RNA as a template for reverse transcription reaction, the operation steps are as follows:

[0048] ① Prepare the following mixture in a PCR tube, and the reaction system is as follows:

[0049] Template RNA (300ng / μL) 2.0μL

[0050] Oligo(dT)18 Primer (50μM) 2.0μL

[0051] RNase free H2O 10.0 μL

[0052] Total 14.0μL

[0053] ② Gently mix and centrifuge, react at 70°C for 10 minutes, then qu...

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Abstract

The invention relates to a method for preparing IKU2 gene of peony purpura, belonging to the technical field of bioengineering. The preparation method of the gene is as follows: (1) extraction of total RNA from the seeds of P. purpurea, (2) synthesis of the first strand of cDNA, (3) amplification of the conserved region of the IKU2 gene of P. purpura, (4) recovery of PCR amplification products Purification, (5) ligation of target fragments and cloning vector pMD 19‑T, (6) preparation of E. coli DH5α strain competent cells, (7) transformation of ligation products, (8) extraction of recombinant plasmids, (9) sequencing. According to the invention, the gene IKU2 regulating seed size is screened from transcriptome libraries at different development stages of the peony seeds, and molecular basis is provided for improving the yield of the peony seeds through genetic engineering technology.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for preparing IKU2 gene of peony purpura. Background technique [0002] Peony is a perennial deciduous shrub of the genus Oaeonia in the family Paeoniaceae. The development and utilization of woody oil can effectively alleviate the shortage of edible oil in my country. In March 2011, the National Ministry of Health Supervision Bureau publicly approved peony seed oil as a new Resource food, peony seed oil is designated as a new type of healthy woody edible oil. At present, the peony species for oil used in large-scale demonstration planting in China mainly include Fengdan (P.ostii) suitable for the southern climate and purple spot peony (P.suffruticosaAndr.var.papaveracea) suitable for the northern climate. But its small seeds (0.88±0.04cm in transverse diameter and 1.00±0.07cm in longitudinal diameter) restrict the efficient development and utilizatio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10A01H5/10A01H6/00
CPCC07K14/415C12N15/8267
Inventor 丁健韩平阮成江闫蕊刘凌悦吴波董旭
Owner DALIAN NATIONALITIES UNIVERSITY
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