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Artificial synthesized Bt insecticidal gene for transgenic anti-insect plants

A gene and gene sequence technology, applied in the field of biological control, can solve problems such as failure to achieve plant protection and inability to express genes efficiently.

Active Publication Date: 2009-11-18
CHINA AGRI UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

[0007] In our laboratory, the Cry1Ac gene has been modified by gene coding frame length and codon optimization. Cry1Ac is highly toxic to lepidopteran insects (such as corn borer), but the original gene cannot be expressed efficiently in plants and cannot reach the target level of plants. According to the requirements of protection, we carried out a comprehensive transformation of the Cry1Ac gene according to the coding characteristics of monocotyledonous plants, and the transformed Cry1Ac-M was connected to the expression vector with CaMV35S-Adh1 as the promoter

Method used

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  • Artificial synthesized Bt insecticidal gene for transgenic anti-insect plants
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  • Artificial synthesized Bt insecticidal gene for transgenic anti-insect plants

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Embodiment Construction

[0033] 1. Modification of Cry1Ac gene

[0034] The present invention carefully analyzes the active region of the Cry1Ac protein, and on the premise of further improving its expression level, transforms the gene coding frame and codons of the Cry1Ac according to the coding characteristics of monocotyledonous plants. The modified Cry1Ac-M has only 69% sequence homology with Cry1Ac, the length of the coding frame has changed from the original 1179 amino acids to 616 amino acids, and the G+C content has also changed from the original 37.32% to 57.64%. See the table below for sub usage. The sequence before the codon change is shown in SEQ ID NO5, and the sequence after the modification is shown in SEQ ID NO6.

[0035]

[0036]

[0037]

[0038]

[0039] 2 Construction of expression vector of Bt Cry1Ac-M gene

[0040] Insertion sequence size and structure, analytical methods to determine its properties

[0041] The size of the inserted sequence is about 3234bp, as follo...

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Abstract

The invention discloses an artificial synthesized Bt insecticidal gene for transgenic anti-insect plants, further, the invention relates to the optimization and transformation systematically to a coding frame and a codon of a CrylAc anti-insect gene, according to an activity analysis to Bt protein in different regions and in accordance with the coding characteristics of the monocotyledons, obtaining anti-insect gene CrylAc-M which can be expressed in a monocotyledons efficiently, with the optimized codon and coding frame. The CrylAc-M gene after being transformed is connected on an expression carrier with CaMV35S-Adhl as promoter, and then the Bt gene CrylAc-Adhl is transferred into a maize inbred line with highly efficient conversion rate by a gene gun co-transformation method. A transgenic progeny is positive through Bt protein testing and a field anti-insect test of the field is tested to be highly resistant.

Description

Technical field: [0001] The invention belongs to the technical field of biological control, and in particular relates to the Bt Cry1Ac-M gene used for plant expression. Background technique: [0002] The Bt gene encodes an insecticidal crystal protein, which is derived from Bacillus thuringiensis, which is a Gram-positive Agrobacterium belonging to the genus Bacillus. It produces insecticidal parasporal crystal proteins called delta-endotoxins during sporulation (the gene that controls the synthesis of this protein is on a plasmid), and these proteins are harmful to Lepidoptera, Diptera, Coleoptera, Hymenoptera and other insects have specific insecticidal activity. [0003] In 1981, Schenpf and Whiteley cloned the first cry gene Cry1Aa1 encoding δ-endotoxin from Bacillus thuringiensis (Bacillus thuringiensis). In 1985, Adang, M.J et al. cloned Cry1Ac gene from Bacillus thuringiensis (Bacillus thuringiensis), but Due to the characteristics of this gene sequence, it has not ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/32C12N15/63C12N15/74C12N15/82C12R1/07
Inventor 赖锦盛董永彬宋伟彬赵海铭
Owner CHINA AGRI UNIV
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