A kind of ginkgo gbflsa gene and its expression protein and application

A ginkgo, gene technology, applied in the field of ginkgo GbFLSa gene and its expression protein and application, can solve the problem of the function research in plants without key enzyme genes, etc.

Active Publication Date: 2022-02-01
NANJING FORESTRY UNIV
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Problems solved by technology

[0004] At present, FLS has been confirmed in model plants to be related to the accumulation of flavonoids in vivo. However, only prokaryotic expression and in vitro activity detection of GbFLS have been carried out in Ginkgo biloba. Research

Method used

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  • A kind of ginkgo gbflsa gene and its expression protein and application
  • A kind of ginkgo gbflsa gene and its expression protein and application
  • A kind of ginkgo gbflsa gene and its expression protein and application

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Embodiment 1

[0026] 1. Cloning of GbFLSa gene by RACE technology

[0027] Ginkgo DNA was extracted using the Plant Genomic DNA Kit (cetyltrimethylammonium bromide) (Zoman, Beijing, China). Total RNA was extracted from Ginkgo biloba leaves using RNAprep Pure Plant kit (Polysaccharides&Polyphenolics-rich, TIANGEN, Beijing, China). Based on Ginkgo biloba transcriptome data (NCBI Short Reads Archive database under access number SRP137637), specific primers for GbFLSa gene were designed, and the full-length cDNA sequence of GbFLSa was cloned by rapid amplification of cDNA ends (RACE). Nested primers were designed to amplify full-length cDNA using the SMATer RACE 5' / 3' Kit (Clontech, Palo Alto, CA, USA) kit. All primers were designed using Oligo 6.0 software (Table 1). It was amplified by PCR, recovered and purified by gel cutting, and transformed into E. coli competent cells through pMD19-T vector. Colonies were detected by PCR, and positive colonies were selected for Sanger sequencing. The...

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Abstract

The invention discloses a Ginkgo biloba GbFLSa gene, its expressed protein and its application, and belongs to the technical field of plant genetic engineering. The nucleotide sequence of the GbFLSa gene of Ginkgo biloba is shown in SEQ ID No.1, and the amino acid sequence of its expressed protein is shown in SEQ ID No.3. By transgenicing the GbFLSa gene of Ginkgo biloba into Populus japonicus, it was found that catechin in the transgenic poplar The content of proanthocyanidins such as procyanidin, epicatechin, epigallocatechin, and gallocatechin was significantly reduced. In addition, the expression levels of DFR, ANS, and LAR enzyme genes were also significantly lower than those of the control. It indicated that GbFLSa encoded a functional protein, which negatively regulated the biosynthesis of proanthocyanidins. It helps to reveal the role of GbFLSa in plant metabolism and better understand the underlying molecular mechanism of flavonoid biosynthesis.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and more specifically relates to a ginkgo GbFLSa gene and its expressed protein and application. Background technique [0002] Flavonoids are widely distributed in the plant kingdom, which can be divided into different subfamilies according to the degree of C epoxidation and saturation, and are one of the representative important secondary products. The rich variety of flavonoids are all composed of 15 carbon atoms arranged in C 6 -C 3 -C 6 The configuration, that is, two aromatic rings in the middle and a ring-forming C 3 link up. It is well known that the biosynthesis of plant flavonoids is a complex process involving many important enzymes. Its synthesis uses coumaryl-CoA and malonyl-CoA as precursors to form yellow chalcone under the action of chalcone synthase (CHS), which is the first limit of the flavonoid synthesis pathway. Quick steps. Chalcone isomerase (CHI) ca...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/53C12N9/02C12N15/82A01H5/00A01H6/00
CPCC12N9/0004C12N15/825
Inventor 徐立安吴雅琼辛月周鹏燕
Owner NANJING FORESTRY UNIV
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