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Coleoptera pest efficient Bacillus thuringiensis cry8G gene, protein and uses thereof

A technology of Bacillus aureus and Coleoptera, which is applied in the technical field of biological control and can solve problems such as difficult to achieve continuous control

Active Publication Date: 2008-01-30
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI +1
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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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  • Coleoptera pest efficient Bacillus thuringiensis cry8G gene, protein and uses thereof
  • Coleoptera pest efficient Bacillus thuringiensis cry8G gene, protein and uses thereof
  • Coleoptera pest efficient Bacillus thuringiensis cry8G gene, protein and uses thereof

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Experimental program
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Effect test

Embodiment 1

[0069] 1.1 Identification of cry gene in strain 185

[0070] A pair of universal primers were designed according to the conserved regions of cry8 genes

[0071] SN5un8 5`GTCCGAATAATCAGAATGAATATG-3`

[0072] SN3un8 5`-CGTTTCGCCTCTCTCACTGCAT-3`

[0073] 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.

[0074] Table 2 The pairing situation of the primers and the conserved regions of cry8 genes and the position of the paired regions on the gene

[0075] Gene

[0076] Cry8Bc

[0077] Table 3 PCR amplification products and restriction enzyme length polymorphisms of cry3

[0078]

[0079] Bt strain 185 was identified using the following PCR reaction (50 μL):

[0080] 10×PCR buffer 5μL

[0081] MgCl 2 (20mM) 6μL

[0082] dNTP (10mM) 1μL

[008...

Embodiment 2

[0116] 2.1 Nucleotide sequence of artificially designed and synthesized cry8G gene that can be expressed in plants

[0117] According to the difference in codon preference between microorganisms and plants, the sequence of 1-2040bp of cry8Gal gene is optimized. The present invention carries out the whole gene synthesis according to the artificially modified sequence of the cry8Gal gene, and the new gene is shown in the nucleotide sequence shown in SEQ ID NO 3. The nucleotide sequence homology between the cry8Gal gene and the mcry8Gal (modified cry8Cal) gene was only 86.88%, and the G+C content was also increased from 37.6% of the original cry8Cal to 45.2% (Table 8). The amino acid codon usage frequency of the Cry8Gal protein was adjusted so that the amino acid codon usage frequency of the mCry8Gal protein was close to the usage frequency in plants (Table 9). The two ends of the artificially modified sequence of the cry8Ca gene were introduced into BamHI and KpnI sites, connec...

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Abstract

The invention relates to a highly effective bacillus thuringiensis cry8G gene and albumen for killing coleoptera pests and application thereof, pertaining to biological control technical field. The invention provides nucleotide sequences of the bacillus thuringiensis cry8G gene which is high poisonous to the coleoptera pests and amino acid sequences of the codes of the protein, and provides nucleotide sequences of the bacillus thuringiensis cry8G gene which is designed artificially and used for transgenic plants and amino acid sequences of the codes thereof. Therefore, toxicity to related pests is presented by the sequences through the genes or the artificial designed sequences of transgenic microbes and plants so as to overcome or delay the generation of drug-resistance of the pests for engineering bacteria and the transgenic plants.

Description

technical field [0001] The invention belongs to the technical field of biological control. The invention relates to the nucleotide sequence of the cry8G gene highly toxic to the Coleopteran pests, to the amino acid sequence of the protein highly toxic to the Coleopteran pests, and to artificially designed and synthesized cry8G genes that can be used in plants. The nucleotide sequence of the cry8G gene expressed in , relates to the amino acid sequence of the protein encoded by the nucleotide sequence of the artificially designed cry8G gene, relates to a recombinant strain containing the cry8G 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 clas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/31C07K14/32C12N15/82A01N63/02C12R1/07A01N63/23A01N63/50
CPCC12R1/075A01N63/02C07K14/325C12N15/8286Y02A40/146A01N63/50A01N63/23C12R2001/075C12N1/205
Inventor 束长龙宋福平冯书亮张杰王容燕黄大昉郎志宏
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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