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Ginkgo GbFLSa gene and expression protein and application thereof

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

Active Publication Date: 2020-06-09
NANJING FORESTRY UNIV
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  • Abstract
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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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  • Ginkgo GbFLSa gene and expression protein and application thereof
  • Ginkgo GbFLSa gene and expression protein and application thereof
  • Ginkgo GbFLSa gene and expression protein and application thereof

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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 GbFLSa gene and an expression protein and application thereof, and belongs to the technical field of plant genetic engineering. The nucleotide sequence of the ginkgo GbFLSa gene is as shown in SEQID No.1, and the amino acid sequence of the expression protein is SEQID No.3. The ginkgo GbFLSa gene is subjected to transgenosis into Populus davidiana x P.bolleana, theinventor finds that the content of proanthocyanidins including catechin, epicatechin, epigallocatechin, gallocatechin and the like in transgenic poplar is notably reduced, and in addition, the expression level of DFR, ANS and LAR enzyme genes is also notably lower than that of the control group, which indicates that GbFLSa coding protein is a functional protein, and has the effect of negative adjustment and control on biologic synthesis of the proanthocyanidins. Disclosure of the effect of the GbFLSa to plant metabolism is facilitated, and the potential molecular mechanism of flavonoid biologic synthesis can be better known.

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