Synthetic genes encoding CRY1AC

A coding, nucleotide sequence technology, applied in genetic engineering, recombinant DNA technology, peptides, etc., can solve problems such as insufficient control of heliothine

Inactive Publication Date: 2010-09-01
BASF AGRICULTURAL SOLUTIONS SEED LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some heliothines cannot be adequately controlled using this technique (Bacheler and Mott (1997) In: Dugger P, Richter D, eds. Beltwide Cotton Conference Proceedings, pp. 858-861. Memphis: National Cotton Council; Smith (1998) In: Dugger P , Richter D, Ed. Beltwide Cotton Conference Proceedings, pp. 965-966. Memphis: National Cotton Council), and this difference was attributed to different Cry 1Ac levels

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Example 1. Synthetic Nucleotide Sequence Encoding Cry1Ac

[0090] A synthetic nucleotide sequence (SEQ ID NO: 6) encoding Cry1Ac was designed. These sequences are represented herein by SEQ ID NO: 1 (synFLCry1Ac), SEQ ID NO: 2 (synCry1Ac-variant 1), SEQ ID NO: 3 (synCry1AcB), SEQ ID NO: 4 (synCry1AcC) and SEQ ID NO: 5 ( optCry1Acv02) said.

Embodiment 2

[0091] Example 2. Expression of synthetic cry1Ac sequences in Bacillus

[0092] The synthetic cry1Ac sequence was amplified by PCR and cloned into the Bacillus expression vector pAX916 by methods well known in the art. After transformation into cells of the cry(-) Bacillus thuringiensis strain, the resulting clones were assayed for expression of the Cry1Ac protein. Bacillus strains comprising the synCry1Ac clone and expressing the Cry1Ac insecticidal protein are grown, for example, in CYS medium (10 g / l Bacto-casitone; 3 g / l yeast extract; 6 g / l KH 2 PO 4 ;14g / l K 2 HPO 4 ; 0.5mM MgSO 4 ; 0.05mM MnCl 2 ;0.05mM FeSO 4 ) until sporulation was evident by microscopic examination. Samples were prepared and analyzed by polyacrylamide gel electrophoresis (PAGE).

Embodiment 3

[0093] Example 3. Determination of insecticidal activity

[0094] The ability of the synthetic cry1Ac nucleotide sequence of the present invention to produce an insecticidal protein was detected. The ability of pesticidal proteins to act as pesticides against pests is often assessed in a number of ways. One method known in the art is to perform feeding assays. In this feeding assay, pests are exposed to a sample containing the compound to be tested or to a control sample. Typically, this is done by placing the material to be tested, or a suitable dilution of such material, on a substance that the pest will ingest, such as an artificial diet. The material to be tested can consist of liquids, solids or slurries. The material to be tested can be placed on the surface and allowed to dry. Alternatively, the material to be tested can be mixed with a molten artificial diet and dispensed into the assay chamber. Assay chambers can be, for example, cups, dishes, or wells of a mic...

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PUM

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Abstract

Compositions and methods for conferring pesticidal activity to bacteria, plants, plant cells, tissues and seeds are provided. Compositions containing a synthetic nucleotide sequence encoding a Cryl Ac protein are provided. The coding sequences can be used in DNA constructs or expression cassettes for transformation and expression in plants and bacteria. Compositions also include transformed bacteria, plants, plant cells, tissues, and seeds. In particular, isolated pesticidal nucleic acid molecules are provided, wherein the nucleotide sequences are set forth in SEQ ID NO:5, 1, 2, 3, or 4, as well as variants and fragments thereof.

Description

field of invention [0001] The present invention relates to the field of molecular biology. Novel nucleotide sequences encoding pesticidal proteins are provided. These proteins and nucleic acid sequences encoding them can be used in the preparation of insecticides and in the production of transgenic pest-resistant plants. Background of the invention [0002] Bacillus thuringiensis is a spore-forming, Gram-positive soil bacterium characterized by its ability to produce insecticides that are specifically toxic to certain orders and species of insects, but are harmless to plants and other non-target organisms. Crystalline inclusions. Accordingly, compositions comprising Bacillus thuringiensis strains or their insecticidal proteins are useful as environmentally acceptable insecticides for the control of agricultural insect pests, or as insect vectors against a variety of human or animal diseases. [0003] The utilization of commercial, transgenic crops expressing Bacillus thur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82
CPCC07K14/325C12N15/8286Y02A40/146
Inventor N·卡罗兹N·德赛D·J·汤姆索
Owner BASF AGRICULTURAL SOLUTIONS SEED LLC
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