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Bt protein expression system

An expression system and bt protein technology, applied in the biological field, can solve the problems of low content of Cry1Ab/Ac and difficulty in purification

Active Publication Date: 2016-02-03
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These safety evaluations require a large amount of high-purity target protein, but the content of Cry1Ab / Ac expressed in rice species is extremely low, and it does not have a label, so it is difficult to purify

Method used

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Experimental program
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Embodiment 1

[0028] 1.1 Experimental material strains and plasmids

[0029] The strains and plasmids used in this paper are shown in (Table 1), and can be distributed to the public.

[0030] Table 1 Strains and plasmids

[0031]

[0032] 1.2 Reagents

[0033] 1.2.1 Medium

[0034] Liquid LB: Tryptone 10.0g, Yeastextract 5.0g, NaCl 10.0g, add water to 1000mL, adjust pH to 7.0, 15 pounds for 20 minutes. Solid LB: Add 1.3% agar to the liquid medium. 1 / 2 solid LB: Tryptone 5.0g, Yeastextract 2.5g, NaCl 5.0g, add water to make up to 1000mL, add 1.3% agar powder, adjust pH to 7.0, 15 pounds for 20 minutes.

[0035] 1.2.2 Antibiotics

[0036] Ampicillin, prepared as a 100 mg / mL aqueous solution, sterilized by filtration with a disposable filter of 0.22 μm, and stored at -20°C. Chloramphenicol and erythromycin were dissolved in absolute ethanol to prepare a 34mg / mL solution and stored at -20°C.

[0037] 1.2.3 Chemical reagents and enzyme reagents

[0038] Restriction enzymes, 2× ligatio...

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Abstract

The invention relates to a Bt protein expression system, belonging to the technical field of biology. A CrylAb / Ac fusion protein expression carrier is characterized in that a carrier pHT315 is inserted by an mcrylAb / ac-C2 sequence, a promoter thereof is a crylAc promoter, and the mcrylAb / ac-C2 sequence is shown as SEQ ID NO: 6. The Bt protein expression system established accordingly can obtain a great amount of high-purity CrylAb / Ac fusion protein, and a foundation is laid for the safety evaluation of transgenic rice such as Huahui No. 1 and the preparation of a CrylAb / Ac protein standard substance.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a Bt protein expression system for expressing Cry1Ab / Ac fusion protein. Background technique [0002] Rice is one of the main food crops in my country. Genetically modified rice mainly includes insect-resistant, disease-resistant, herbicide-resistant, stress-resistant environments, and improved rice quality. Transgenic cry1Ab / Ac rice (Huahui No. 1) is a rice variety independently bred by Huazhong Agricultural University with strong insecticidal activity against rice stem borer, stem borer, and rice leaf roller (Field performance of transgenicelite commercial hybrid rice expressing Bacillus thuringiensis delta-endotoxin.TuJM, et al. .NatureBiotechnology,2000,18(10):1101-1104), in 2009, the Bt insect-resistant rice Huahui 1 and Shanyou 63 obtained the biosafety certificate (The first approved transgenic rice in China.LuC.GMCrops,2010,1(3):113- 115), has broad application prospects. H...

Claims

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Application Information

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IPC IPC(8): C12N15/75C12N1/21C12P21/02C07K19/00C07K1/18C07K1/16C12R1/07
Inventor 张杰耿丽丽束长龙彭琦宋福平梁影屏
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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