Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Insecticidal gene and use thereof

A technology of insecticidal genes and transgenics, applied in the direction of genetic engineering, plant genetic improvement, application, etc., can solve problems such as environmental pollution, increased production costs, and the need for repeated application

Inactive Publication Date: 2013-06-12
BEIJING DABEINONG TECH GRP CO LTD +1
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002]Pest pests are the main factor leading to crop loss, causing significant economic losses to farmers, and even affecting the survival of the local population
In order to control plant pests, people usually use broad-spectrum chemical insecticides and biological insecticides, but both have limitations in practical application: chemical insecticides will cause environmental pollution problems and lead to the emergence of resistant insects. However, biopesticides are easily degraded in the environment and need to be repeatedly applied in production, which greatly increases the production cost

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Insecticidal gene and use thereof
  • Insecticidal gene and use thereof
  • Insecticidal gene and use thereof

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0067] The first embodiment, selection, optimization and synthesis of PIC2 gene

[0068] 1. Select the PIC2-01 gene sequence for optimization

[0069] The amino acid sequence of the natural Cry1Ba3 gene whose serial number was AAK63251.1 registered by Zhang Jie et al. in 2003 is the basis for the optimization of the insecticidal gene PIC2-01 of the present invention. The nucleotide sequence of the natural Cry1Ba3 gene is as shown in SEQ ID NO in the sequence table 2, the amino acid sequence of the natural Cry1Ba3 gene is shown in SEQ ID NO: 3 in the sequence listing.

[0070] 2. Optimizing the amino acid sequence of the above-mentioned PIC2-01

[0071] Keep the above PIC2-01 amino acid sequence (1228 amino acids) unchanged, and optimize the nucleotide sequence (3687 nucleotides) encoding the above PIC2-01 amino acid sequence (1228 amino acids) according to the corn preference codon remodel.

[0072] The strategies of codon optimization and transformation mainly include: acc...

no. 2 example

[0076] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0077] 1. Construct the recombinant cloning vector DBN01Ba-T containing the PIC2-01 nucleotide sequence and connect the synthetic PIC2-01 nucleotide sequence into the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), the operation steps According to the instructions of the pGEM-T vector of Promega company, the recombinant cloning vector DBN01Ba-T was obtained, and its construction process was as follows: figure 1 Shown (wherein, Amp represents the ampicillin resistance gene; f1 represents the replication origin of phage f1; LacZ is the LacZ start codon; SP6 is the promoter of SP6 RNA polymerase; T7 is the promoter of T7 RNA polymerase; PIC2-01 is the promoter of PIC2 -01 nucleotide sequence (SEQ ID NO: 1); MCS is a multiple cloning site).

[0078] Then, the recombinant cloning vector DBN01Ba-T was transformed into Escher...

no. 3 example

[0088] The third embodiment, the acquisition and verification of corn plants transferred to PIC2-01 nucleotide sequence

[0089] 1. Obtaining maize plants transferred to the PIC2-01 gene sequence

[0090] According to the routinely adopted Agrobacterium infection method, the immature embryos of the aseptically cultured corn variety Zong 31 (Z31) were co-cultured with the Agrobacterium described in 4 in the second embodiment, so that 2 and 3 in the second embodiment The T-DNA in the constructed recombinant expression vectors DBN100056 and DBN100056N (native sequence) (including the promoter sequence of maize Ubiquitin gene, PIC2-01 nucleotide sequence, native sequence, PMI gene and Nos terminator sequence) were transferred to maize In the genome, maize plants transformed with PIC2-01 nucleotide sequence and maize plants transformed with natural sequence (positive control) were obtained; meanwhile, wild-type maize plants were used as negative control.

[0091] For Agrobacterium...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to an insecticidal gene and use thereof. The nucleotide sequence of the insecticidal gene comprises (a) a nucleotide sequence shown as SEQ ID NO:1; or (b) a sequence in the same code with the nucleotide sequence shown in (a) and is not SEQ ID NO:2; or (c) a nucleotide sequence which is hybridized with the nucleotide sequence limited by (a) or (b) under strict conditions and codes proteins with insecticidal activity. The insecticidal gene provided by the invention is in particular suitable for being expressed in monocotyledon, in particular in corn and paddy, so that not only is the expression quantity and stability of PIC2-01 insecticidal gene remarkably improved, but also the toxicity of the PIC2-01 insecticidal gene to injurious insects is further remarkably enhanced, in particular to lepidopteron.

Description

technical field [0001] The invention relates to an insecticidal gene and its application, in particular to a modified PIC2-01 insecticidal gene and its application. Background technique [0002] Plant pests are the main factor leading to the loss of crops, causing significant economic losses to farmers, and even affecting the living conditions of the local population. In order to control plant pests, people usually use broad-spectrum chemical insecticides and biological insecticides, but both have limitations in practical application: chemical insecticides will cause environmental pollution problems and lead to the emergence of resistant insects. However, biopesticides are easily degraded in the environment and require repeated application in production, which greatly increases production costs. [0003] In order to solve the limitations in the practical application of chemical insecticides and biological insecticides, scientists have discovered through research that by tra...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N15/32C12N15/63C12N5/10C12N1/13C12N1/19C12N1/21C12N7/01C12P21/02A01K67/033A01H5/00
Inventor 丁德荣
Owner BEIJING DABEINONG TECH GRP CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com