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Molecular cloning and identifying method for silkworm gene

A technology of molecular cloning and identification method, applied in the field of biology, can solve the problems of difficult decomposition and excretion, greater harm of pesticides, enrichment and other problems

Active Publication Date: 2017-11-24
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] To sum up, the problems in the prior art are: the current pesticides are more harmful to humans and animals, difficult to decompose and excrete, and easy to accumulate in organisms

Method used

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  • Molecular cloning and identifying method for silkworm gene
  • Molecular cloning and identifying method for silkworm gene
  • Molecular cloning and identifying method for silkworm gene

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Embodiment Construction

[0035] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0036] The application principle of the present invention will be described in detail below in conjunction with the accompanying drawings.

[0037] The silkworm gene provided by the embodiment of the present invention is SEQ ID NO: 1; named BmDHODH.

[0038] like figure 1 As shown, the molecular cloning and identification method of the silkworm gene provided by the embodiments of the present invention comprises the following steps:

[0039] S101: RNA extraction, using TRIZOL reagent to extract total RNA. After digesting residual genomic DNA with RNase-free DNase I for 30 min at 37 °C, synthesize first-strand cDNA transcr...

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Abstract

The invention belongs to the field of biology and discloses a molecular cloning and identifying method for a silkworm gene. The sequence of the silkworm gene is SEQ ID NO:1; and the silkworm gene is named BmDHODH. The method disclosed by the invention has the advantages that leflunomide is used for treating silkworm cells and has great inhibitory effect on BmDHODH expression and cell proliferation; as a component in pyrimidine synthesis, BmDHODH is a necessary biological enzyme closely related with cell growth and proliferation, can become a new gene target for killing lepidoptera pests, provides new opinion for eco-friendly pest control and provides a new target and a new theoretical method for developing an environment-friendly pesticide; and as a biological model for the lepidoptera pests, basic knowledge of the lepidoptera pests can be enriched for silkworm BmDHODH, and a new method is provided for environment-friendly pest control.

Description

technical field [0001] The invention belongs to the field of biology, in particular to a method for molecular cloning and identification of silkworm genes. Background technique [0002] As a typical representative of Lepidoptera insects and an ideal biological model, the silkworm has played an important role in promoting the development of animal science and is also an important model for eco-friendly pest control. BmDHODH is a dihydroorotate dehydrogenase that can oxidize dihydroorotate to orotate, the fourth enzyme of pyrimidines in the de novo pyrimidine biosynthesis pathway. First, we cloned and studied the function of BmDHODH in silkworm and found that it was ubiquitously expressed throughout development and in nine different tissues. Then, knockdown of BmDHODH at mRNA and protein levels by leflunomide treatment or dsRNA interference in silkworm cells significantly reduced cell growth and proliferation by inducing cell cycle arrest at G2 / M phase. As a component in pyr...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/10C12Q1/68
CPCC12N9/001C12N15/10C12Q1/6851C12Y103/01014C12Q2531/113C12Q2561/113
Inventor 崔红娟赵二虎蒋小兰张奎杨丽群
Owner SOUTHWEST UNIVERSITY
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