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Efficient bacillus thuringiensis cry8H gene, protein for vaginata destructive insect and uses of the same

A technology of Bacillus aureus and Coleoptera, which is applied in the field of biological control technology and can solve problems such as difficulty in achieving continuous control.

Active Publication Date: 2008-02-27
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to control the harm of grubs, comprehensive control strategies such as agriculture, chemistry, and physics are generally adopted. Although this has certain effects, it is difficult to achieve the effect of continuous control

Method used

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  • Efficient bacillus thuringiensis cry8H gene, protein for vaginata destructive insect and uses of the same
  • Efficient bacillus thuringiensis cry8H gene, protein for vaginata destructive insect and uses of the same

Examples

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Effect test

Embodiment 1

[0067] 1.1 Identification of cry gene in strain BT-SU4

[0068] A pair of universal primers were designed according to the conserved regions of cry8 genes

[0069] SN5un85`-GTCCGAATAATCAGAATGAATATG-3`

[0070] SN3un85`-CGTTTCGCCTCTCTCACTGCAT-3`

[0071] Table 2 shows the homologous sequences of these genes and primers, and Table 3 shows the size of the digested fragments of cry8 gene amplified products predicted by this pair of primers. These genes can be identified respectively by this PCR-RFLP method.

[0072] Table 2 The pairing situation of the primers and the conserved regions of each gene of cry8 and the position of the paired regions on the gene

[0073] Gene

[0074] Table 3 PCR amplification products and restriction enzyme length polymorphisms of cry8

[0075] genotype

[0076] Bt strain 185 was identified using the following PCR reaction (50 μL):

[0077] 10×PCR buffer 5μL

[0078] MgCl 2 (20mM) 6μL

[0079] dNTP (10mM) 1μL

[0080] Prime...

Embodiment 2

[0110] 2.1 Nucleotide sequence of artificially designed and synthesized cry8H gene that can be expressed in plants

[0111] According to the difference in codon preference between microorganisms and plants, the sequence of 1-2010bp of cry8Ha1 gene was optimized. The present invention carries out whole gene synthesis according to the artificially modified sequence of cry8Ha1 gene, and the new gene is shown in the nucleotide sequence shown in SEQ ID NO3. The nucleotide sequence homology between the cry8Ha1 gene and the mcry8Ha1 (modified cry8Ha1) gene was only 87.08%, and the G+C content was also increased from 37.6% of the original cry8Ha1 to 50.1% (Table 8). The amino acid codon usage frequency of the Cry8Ha1 protein was adjusted so that the amino acid codon usage frequency of the mCry8Ha1 protein was close to the usage frequency in plants (Table 9). The two ends of the artificially modified sequence of the cry8Ha gene were introduced into the BamHI and KpnI sites, connected ...

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Abstract

The invention discloses a Bacillus thuringiensis cry8H gene, protein and application to kill Coleoptera pest in the biological prevention technical domain, wherein the nucleotide sequence of high-toxicity cry8H gene of Coleoptera pest is displayed as SEQ ID NO1; the nucleotide sequence of the expressed cry8H gene synthesized by artificial design is displayed as SEQ ID NO3, which has the property to resist Holotrichia oblita, Anomala corpulenta and Holotrichia parallela.

Description

technical field [0001] The invention belongs to the technical field of biological control. The invention relates to the nucleotide sequence of cry8H gene highly toxic to coleopteran pests, to the amino acid sequence of a protein highly toxic to coleopteran pests, to artificially designed and synthesized cry8H genes that can be used in plants The nucleotide sequence of the cry8H gene expressed in , relates to the amino acid sequence of the protein encoded by the nucleotide sequence of the artificially designed cry8H gene, relates to a recombinant strain containing the cry8H gene, relates to the use of the gene to construct an expression vector, and also relates to the use of the above-mentioned gene Sequences for methods of plant transformation. Background technique [0002] Scarab belongs to Coleoptera Scarabaeidae (Scarabaeidae), and its larvae (commonly known as grubs, also referred to as "grubs" for short in the present invention hereinafter) are a class of important worl...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N15/31C07K14/32A01N63/02C07H21/04C12N15/82A01P7/04A01N63/23A01N63/50
CPCC12R1/075C12N15/8286C07K14/325A01N63/02A01N63/00Y02A40/146A01N63/50A01N63/23C12R2001/075C12N1/205
Inventor 束长龙宋福平王容燕张杰冯书亮黄大昉郎志宏
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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