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Preparation method of panonychus citri me gregor chitin gene-based double-target transgenetic vector

A technology of transgenic vector and chitinase gene, applied in the field of transgenic to achieve the effect of reducing usage and broad market prospects

Inactive Publication Date: 2013-04-03
CITRUS RES INST SOUTHWEST UNIV
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Problems solved by technology

[0003] The purpose of the embodiment of the present invention is to provide the preparation and application of a dual-target transgenic vector based on the chitin gene of Panonychus citrus, aiming to solve the problem of using RNA interference technology to obtain insect-resistant materials

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  • Preparation method of panonychus citri me gregor chitin gene-based double-target transgenetic vector

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Embodiment Construction

[0011] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0012] figure 1 A flowchart showing the preparation method of the dual-target transgenic vector based on the chitin gene of Panonychus citrus provided by the embodiment of the present invention.

[0013] In this preparation method, two chitinase genes, Unigene10731 and Unigene7021, were identified from the transcriptome database of Panonychus citrus. After the authenticity test, the two selected genes were compared on NCBI to find out the conserved sequence. According to the multigene large fragment construction strategy, two genes were spliced ​​into a multigene fragment, and Unigene10731 and Uni...

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Abstract

The invention discloses an application method of 2 chitinase genes. Two chitinase genes Unigene10731 and Unigene 7021 are identified from an orange panonychus citri me gregor transcriptome database and are subjected to authenticity testing; the two selected genes are compared on NCBI (National Center for Biotechnology Information) to find a conserved sequence; and the two genes are jointed into a multi-gene segment according to the multi-gene large-segment construction strategy, and the Unigene10731 and the Unigene 7021 are jointed together to form a multi-gene; a primer containing a restriction enzyme cutting site is designed, and forward and reverse segments of the multi-gene chitinase order are sequentially jointed into two terminals of the intron of the vector, so that an RNAi (ribonucleic acid interfere) interference vector 'PFGC+Ug10731 / Ug7021' with a reverse complementary hairpin structure can be formed, and the vector is reprinted into agrobacterium to infect an orange nursery stock.

Description

technical field [0001] The invention belongs to the field of transgenic technology, and in particular relates to the preparation and application of a double-target transgenic carrier based on the chitin gene of Panonychus citrus. Background technique [0002] Panonychus citrus is a worldwide pest of citrus, which has a great impact on the yield and quality of citrus. At present, chemical control is still an important measure to control Panonychus citrus. Improper application of chemical pesticides not only affects fruit safety, but also leads to serious drug resistance. Obtaining insect-resistant transgenic materials based on RNA interference technology has been successful in crops such as rice, vegetables, and cotton. Insect chitinase is involved in important life activities such as insect (mite) molt, peritrophic membrane degradation and defense. The construction of RNA interference vector through the chitinase gene of Panonychus citrus, and then obtaining transgenic cit...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/113C12N15/66A01H5/00
Inventor 冉春张云飞刘浩强陈飞李鸿筠李俊丽李晓姣岳建书
Owner CITRUS RES INST SOUTHWEST UNIV
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