Bacillus thuringiensis cry1Ah/cry1Ie bivalent gene expression vector and application thereof

A m2-cry1ah, m2-cry1ie technology, applied in the biological field, can solve the problems of low protein expression and low transformation efficiency, and achieve the effect of high expression and good application value

Active Publication Date: 2013-08-28
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Abstract
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Problems solved by technology

However, the biolistic method is used for the transformation of bivalent genes, and the transformation efficiency is not high enough, and the protein expression level in plants is not high. It is necessary to further transform it to meet the needs of large-scale production and breeding.

Method used

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  • Bacillus thuringiensis cry1Ah/cry1Ie bivalent gene expression vector and application thereof
  • Bacillus thuringiensis cry1Ah/cry1Ie bivalent gene expression vector and application thereof
  • Bacillus thuringiensis cry1Ah/cry1Ie bivalent gene expression vector and application thereof

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Embodiment Construction

[0047] The present invention is described in further detail below in conjunction with embodiment.

[0048] The biological materials used below are preserved in the applicant's laboratory and can be distributed to the public.

[0049] 1. Imported genes:

[0050] Target gene: In this study, the cry1Ah and cry1Ie genes were optimized twice, and the modified cry1Ah gene (m2-cry1Ah) was compared with the original cry1Ah gene: the GC content increased from 37% to 55%. The GC content of the optimized cry1Ie gene (m2-cry1Ie) was increased to 55%. See the sequence listing for the modified nucleotide sequence and deduced amino acid sequence. Table 1 shows the GC content and codon usage frequency of the secondary transformed cry1Ah and cry1Ie genes

[0051] Table 1 GC content and codon usage frequency of transformed cry1Ah and cry1Ie genes

[0052]

[0053]

[0054] 2. Construction of recombinant vector pMAhIeb:

[0055] Using pCAMBIA3300 as the vector backbone, the insert seq...

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Abstract

The invention relates to a bacillus thuringiensis cry1Ah / cry1Ie bivalent gene expression vector and an application thereof, belonging to the technical field of biology. pCIABIA3301 is taken as a vector framework, and m2-cry1Ah and m2-cry1Ie genes are inserted on multiple cloning sites of the pCIABIA3301 to get the expression vector pMAhIeb. An agrobacterium-mediated transformation method is utilized for transforming the constructed expression vector to a species of a maize inbred line to get a high-resistance transgenic bivalent gene maize transformation event. The research of expression and genetic stability of a Cry1Ah protein of the two transformation events shows that the Cry1Ah protein can realize normal and stable expression in T1-T2 generations of maize plants; and furthermore, the expression level is very high, and the resistance of the plants to pyrausta nubilalis in a field is also genetic stably. The obtained insect-resistant transgenic maize material has great application value and can be used as a candidate material for next-step Bt insect-resistant maize breeding work.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a bacillus thuringiensis cry1Ah / cry1Ie bivalent gene expression vector and its application in transgenic plants. Background technique [0002] Corn (Zea mays L.) is an important food crop, industrial raw material and feed raw material, and plays a very important role in the development of the national economy. my country's corn production has continued to develop in recent years. In 2012, the planting area reached 510 million mu, and the total output reached 208 million tons, surpassing rice to become my country's largest food crop. However, maize production is also restricted by many factors. In recent years, the severe occurrence of lepidopteran pests such as corn borer and armyworm has become an important factor affecting the increase of maize production. In particular, the corn borer occurs year after year in northern my country, causing an average reduction of about 5% in corn ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84A01H5/00
Inventor 黄大昉郎志宏朱莉李秀影李圣彦戴军汪海
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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