Ginkgo long-chain non-coding RNA and application thereof to bilobalide biosynthesis

A long-chain non-coding, ginkgolide technology, applied in DNA/RNA fragments, applications, recombinant DNA technology, etc., can solve the problem of lncRNA in the blank stage

Pending Publication Date: 2021-12-21
HUNAN UNIV OF SCI & ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, at present, the application of lncRNA in the regulation of ginkgolide biosynthesis in Ginkgo biloba, an important economic tree species, is still in a blank stage.

Method used

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  • Ginkgo long-chain non-coding RNA and application thereof to bilobalide biosynthesis
  • Ginkgo long-chain non-coding RNA and application thereof to bilobalide biosynthesis
  • Ginkgo long-chain non-coding RNA and application thereof to bilobalide biosynthesis

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Experimental program
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Embodiment 1

[0012] 1. Collection of samples

[0013] The 5-year-old healthy male ginkgo tree was used as the test material, and the leaf and root samples (as a control) were collected respectively, immediately put into liquid nitrogen, and stored at -80°C for future use.

[0014] 2. RNA extraction

[0015] RNA was isolated and purified from Ginkgo biloba leaves and roots using Trizol kit (Invitrogen, Carlsbad, CA, USA). RNA purity and integrity were checked using NanoDrop ND-1000 (NanoDrop, Wilmington, DE, USA) and Agilent 2100, respectively.

[0016] 3. RNA library construction and sequencing

[0017] Via Ribo-Zero TM After removal of rRNA by the rRNA Removal Kit (Illumina, San Diego, USA), the remaining RNA was fragmented into small pieces using divalent cations at high temperature. The cleaved RNA fragments were reverse transcribed into cDNA, which was then used to synthesize U-labeled second-strand DNA using E. coli DNA polymerase I, RNase H, and dUTP. A bases are then added to t...

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Abstract

The invention belongs to the technical field of plant molecular biology, and particularly relates to ginkgo long-chain non-coding RNA and an application thereof to bilobalide biosynthesis, and the sequence of the ginkgo long-chain non-coding RNA is shown as SEQ ID NO.1. The ginkgo long-chain non-coding RNA can effectively increase the content of bilobalide.

Description

technical field [0001] The invention belongs to the technical field of plant molecular biology, and particularly relates to ginkgo long-chain non-coding RNA and its application in the biosynthesis of ginkgolide. Background technique [0002] Ginkgo biloba is one of the oldest relict plants with high medicinal and economic value, among which terpene trilactone is a unique component in Ginkgo biloba extract, including diterpene lactone ginkgolide and hemiterpene lactone ginkgo. ester. Ginkgolide is an important active substance in Ginkgo biloba. The main active ingredients can be divided into ginkgolide A, B, C, M and J. It has remarkable effects in the treatment of cardiovascular and neurological diseases and has been widely used in clinical practice. . Due to the particularity and complexity of the chemical structure of ginkgolides, the artificial synthesis process is complicated, the cost of industrialization is high, and the products do not have biological activity. At p...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/82A01H5/00A01H6/00
CPCC12N15/113C12N15/8243C12N2310/10
Inventor 刘晓霞覃佐东罗小芳骆鹰何福林
Owner HUNAN UNIV OF SCI & ENG
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