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Red grape ABA8ox3 gene overexpression vector as well as construction method and application thereof

An overexpression vector and gene overexpression technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problems of grape fruit yield and quality damage, reduced commerciality and other problems, and achieve the effect of reducing the loss of fruit farmers and high commodity value.

Inactive Publication Date: 2021-08-10
ZHEJIANG WANLI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in southern China, due to the influence of the monsoon climate from July to August, the yield and quality of grape fruit were damaged, and the commerciality was reduced

Method used

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  • Red grape ABA8ox3 gene overexpression vector as well as construction method and application thereof
  • Red grape ABA8ox3 gene overexpression vector as well as construction method and application thereof
  • Red grape ABA8ox3 gene overexpression vector as well as construction method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0038] Embodiment 1: Yinhong grape ABA8ox3 gene overexpression vector, it is characterized in that, this overexpression vector is that the purpose fragment of Yinhong grape ABA8ox3 gene is inserted between the CaMV 35 promoter and mgfp5 fluorescent sequence of pCAMBIA1302 original vector and replaces pCAMBIA1302 original The pCAMBIA1302-ABA8ox3 overexpression vector obtained by deleting the "catg" 4 bases of the vector sequence and the stop codon of the target fragment, the nucleotide sequence of the overexpression vector is shown in SEQ ID NO.1.

Embodiment 2

[0039] Embodiment 2: The construction method of the pCAMBIA1302-ABA8ox3 overexpression vector of Yinhong grape ABA8ox3 gene, obtain Yinhong grape ABA8ox3 gene by cloning technology, insert the objective fragment of Yinhong grape ABA8ox3 gene into the designed pCAMBIA1302 original vector. The expressed site realizes the construction of the pCAMBIA1302-ABA8ox3 overexpression vector, which specifically includes the following steps:

[0040] S1, cloning of the coding region of the ABA8ox3 gene of Yinhong grape:

[0041] S1.1, utilize the RNA extraction kit (Tiangen, China) to extract the RNA of red grape pulp, adopt NanoDrop2000 trace nucleic acid protein analyzer (Thermo Fisher, USA) to measure RNA concentration after obtaining RNA, adopt DYCP-31BN type agarose Horizontal electrophoresis instrument was used to analyze the integrity of RNA, and the agarose electrophoresis picture of the ABA8ox3 gene coding region of Yinhong grape was shown in figure 1 ; The extracted RNA was reve...

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Abstract

The invention discloses a red grape ABA8ox3 gene overexpression vector as well as a construction method and an application thereof, the overexpression vector is a pCAMBIA1302-ABA8ox3 overexpression vector obtained by inserting a target fragment of a red grape ABA8ox3 gene between a CaMV 35 promoter and an mgfp5 fluorescent sequence of a pCAMBIA1302 original vector, replacing four catg basic groups of the pCAMBIA1302 original vector sequence and deleting a termination codon of the target fragment, and the nucleotide sequence of the overexpression vector is as shown in SEQ ID NO. 1. The pCAMBIA1302-ABA8ox3 constructed by the invention has the capability of overexpressing the ABA8ox3 gene, and the content of endogenous ABA in red grape fruits is directly regulated and controlled by overexpressing the ABA8ox3 gene, so that the mature period of the red grapes is advanced, the picking period of the red grapes can bypass the typhoon season, the loss of fruit farmers is reduced, and the pCAMBIA1302-ABA8ox3 win the same-period competition with other fruits or grapes and obtains higher commodity value.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an ABA8ox3 gene overexpression vector of Yinhong grape and its construction method and application. Background technique [0002] Grape (scientific name: Vitis vinifera) is one of the five economical fruit trees in my country. Its fruit has rich nutritional value, fresh and sweet unique taste, and various eating methods, and is widely loved by the Chinese people. However, in southern China, due to the influence of the monsoon climate from July to August, the yield and quality of grape fruit were damaged, and the commerciality decreased. The main grape variety "Yinhong" in Ningbo City, Zhejiang Province was affected by this. [0003] The hydroxylation catalyzed by ABA8'-hydroxylase can catabolize ABA (abscisic acid, abscisic acid), and this reaction is encoded by the CYP707A gene family. ABA plays a major regulatory role in the fruit ripening process, so the change of endogenous ABA...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N9/02C12N15/84C12N15/66A01H5/08A01H6/88
CPCC12N9/0073C12N15/8293C12Y114/13093
Inventor 董昳伶吴月燕
Owner ZHEJIANG WANLI UNIV
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