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Strawberry NCED gene and construction of vector thereof

A technology of gene and strawberry, applied in the construction field of strawberry NCED gene and plant expression vector of the gene

Inactive Publication Date: 2011-06-15
CROP RES INST OF FUJIAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are no related reports on strawberry NCED gene

Method used

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  • Strawberry NCED gene and construction of vector thereof
  • Strawberry NCED gene and construction of vector thereof

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Effect test

Embodiment 1

[0014] Embodiment 1 Strawberry NCED gene cloning

[0015] Total RNA was extracted from strawberry fruit using Trizol instructions. The first strand of cDNA was synthesized according to the method of PrimeScriptTM 1st Strand cDNA Synthesis Kit kit from Dalian Bao Biological Co., Ltd. The AP sequence of the reverse transcription primer was 5′-GGCCACGCGTCGACTAGTAC(T)17-3′.

[0016] According to the genebank, soybeans, cowpeas, kidney beans, and radish NCED A pair of degenerate primers for the conserved region were designed for the gene sequence, upstream N1: 5′-GGK CACCACTTCTTCGACGG-3′, downstream N2: 5′-CTCTTCAAACTCTCVTCGCATTC-3′, PCR amplification was performed with these two primers, and a 842 bp gene was amplified fragments such as figure 1 As shown in A.

[0017] under strawberries NCED According to the results of cloning and sequencing of the conserved region of the gene, the 3′-terminal cis primer N3 was designed. The primer sequence is: 5′-ATCTGGGTTGAGTCGCCGG-3′,...

Embodiment 2

[0021] Embodiment 2 Strawberry NCED Gene plant expression vector construction

[0022] under strawberries NCED The coding region of the complete cDNA sequence of the gene, the multiple cloning site of the plant expression vector pBI121, and a pair of specific primers were designed for NCED Amplification of the complete open reading frame of the gene, introduced at the 5' and 3' respectively Xba I and Bam H I restriction site (NCEDX: 5'-GCTCTAGAATGGCTACTCCTTCGAATAGTTG-3'; NCEDB: 5'-CGGGATCC CTATGCCTGGTTTTTCAAGGC-3'), the PCR amplification product was Xba I and Bam H I double digestion, recovery NCED Gene fragment. Plant expression vector pBI121 plasmid Xba I and Bam H I double enzyme digestion, recovery vector fragments. Will NCED The gene fragment and the vector fragment of pBI121 were ligated overnight at 16°C under the action of T4 ligase, and the ligation product was transformed into competent E. coli DH5α, spread on the LB plate containing Kan, and a ...

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Abstract

The invention provides a strawberry NCED gene and a construction of a vector thereof. The nucleotide sequence of the gene is shown in SEQ ID NO:1 or has a cDNA sequence which has more than 90% of homology with the nucleotide sequence shown in SEQ ID NO:1 and is used to code the same functional protein. In the invention, the coding gene of the 9-cis-epoxy carotene dioxygenase is cloned from strawberry, the gene has critical regulating action on the synthesis of ABA in the processes of maturity and wilting or the conditions of drought, high temperature and low temperature. The invention lays the foundation for the research of the strawberry ABA anabolism mechanism and provides a gene source for the genetic engineering regulation of the metabolism and accumulation of ABA in plant.

Description

Technical field [0001] The invention involves the field of genetic engineering, which specifically involves the construction of a strawberry NCED gene and the genetic plant expression carrier. Background technique [0002] As a hormone that plays an important role in the entire life cycle of plants, it has extensive physiological effects in plant embryo development, seed dormant and germination, fruit maturity, and plants' response to adversity coercion.Growth and development plays an important role.The indirect synthesis of ABA needs to be a series of complex reaction changes such as aggregation, circularization, heterogeneity, oxidation, and cracking. Many of the details of the details are not clear. The research on its regulation mechanism has just started.How to express various genes, and the specificity of various enzymes needs to be further confirmed.9-cashmere carotenoids (NCED) catalyzed 9-catalyzed-cashmere-purple yellow and 9-new yellow quality cracking to form yellow a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N15/29C12N9/02C12N15/63C12N15/82A01H5/00
Inventor 朱海生
Owner CROP RES INST OF FUJIAN ACAD OF AGRI SCI
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