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Plant expression vector and application thereof

A plant expression vector and the technology of the vector are applied to the plant expression vector and its application field, which can solve the problems of herbicide resistance and insect resistance cotton need to be improved, and achieve the effects of reducing negative effects and environmental pollution, and reducing human and financial resources.

Active Publication Date: 2013-11-20
XINJIANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the current methods for preparing herbicide-resistant and insect-resistant cotton still need to be improved

Method used

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  • Plant expression vector and application thereof
  • Plant expression vector and application thereof
  • Plant expression vector and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The construction of embodiment 1 plant expression vector

[0043] 1. Reagents and materials:

[0044] The pBI121 empty vector was purchased from Beijing Dingguo Biotechnology Company, plasmid Bt-vip, plasmid Bt-4AB, Agrobacterium tumefaciens engineering strain LBA4404, and Escherichia coli DH5a (all provided by the Key Laboratory of Agricultural Biotechnology of Xinjiang Agricultural University).

[0045] Bar gene primer 1: 5'-ggtctagaatgagcccagaacgacgcc-3' (SEQ ID NO: 2), where tctaga in bold font is the XbaⅠ restriction site;

[0046] Primer 2 of bar gene: 5'-ctcggatcctcagatctcggtgacgggca-3' (SEQ ID NO: 3), where ggatcc in bold font is the BamHI restriction site.

[0047] Bt-4AB Primer 1: 5'-caacggttccgctctttctg-3' (SEQ ID NO: 4),

[0048] Bt-4AB Primer 2: 5'-cgtggttctgcccaaggtat-3' (SEQ ID NO: 5).

[0049] 2. Method

[0050] According to the method for constructing a plant expression vector of the present invention, refer to Figure 1-Figure 3 , construct the pla...

Embodiment 2

[0083] 1. Materials and Reagents

[0084] The embryogenic callus of Xinhai 14 sea-island cotton, which was obtained from hypocotyl-induced callus through somatic embryogenesis, was used as the test material and transformed recipient in this example.

[0085] YEB (liquid / solid) medium: for the cultivation of Agrobacterium tumefaciens; tryptone 5g / L, yeast extract 1g / L, sucrose 5g / L, MgSO 4 ·7H 2 O0.5g / L, agar powder 15g / L (solid), pH=7.0.

[0086] MS co-cultivation medium: glucose 30g / L, coagulant 1.8g / L, MgCl 2 0.75g / L, macroelement I 100ml / L, trace element II 5ml / L, iron salt 5ml / L, B 5 Organic matter 5ml / L, pH=5.8.

[0087] MS 0 Basic culture medium: glucose 30g / L, macroelement I 100ml / L, trace element II 5ml / L, iron salt 5ml / L, B 5 Organic matter 5ml / L, pH=5.8.

[0088] MS 3 Medium (Cotton embryogenic callus transformation medium): glucose 30g / L, coagulant 1.8g / L, MgCl 2 0.75g / L, KNO 3 1.9g / L, Ammonium Asparagine 0.1g / L, Ammonium Glutamine 0.1g / L, Major Element I ...

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Abstract

The invention discloses a plant expression vector and an application thereof. The plant expression vector contains a first expression box, a second expression box and a third expression box, wherein the first expression box contains a first nucleic acid molecule Bt gene, a first promoter (CaMV35S promoter) and a first terminator (NOS terminator); the second expression box contains a second nucleic acid molecule bar gene, a second promoter (CaMV35S promoter) and a second terminator (NOS terminator); and the third expression box contains a third nucleic acid molecule NPT II gene, a third promoter (NOS promoter) and a third terminator (NOS terminator). The plant expression vector is utilized for performing gene transformation, so that an anti-herbicide and anti-pest transgenic plant can be prepared once, the plant resistance can be effectively improved, the production cost is reduced, the yield and the income are increased, and the environmental pollution caused by large-quantity use of pesticides can be also reduced.

Description

technical field [0001] The invention relates to the field of plant transgenic technology. Specifically, the present invention relates to plant expression vectors and applications thereof, more specifically, to plant expression vectors, kits for preparing herbicide-resistant and insect-resistant transgenic plants and their uses, methods for constructing plant expression vectors, and preparation of herbicide-resistant transgenic plants. Agents and methods for insect-resistant transgenic plants. Background technique [0002] Cotton is an important textile fiber crop and an important economic crop. About 180 million people in the world are engaged in activities related to cotton production. my country is the world's largest cotton producer and cotton consumer. Long-term weed damage and pest damage are one of the main factors affecting cotton production. There are many kinds of weeds in cotton fields, with strong adaptability and strong fecundity, which greatly reduces the qual...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/66A01H5/00
Inventor 曲延英陈全家王希东孟灵真孙国清
Owner XINJIANG AGRI UNIV
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