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A method for molecular cloning and identification of silkworm genes

A technology of molecular cloning and identification methods, applied in the field of biology, can solve problems such as difficulty in decomposition and excretion, enrichment, and greater harm of pesticides

Active Publication Date: 2021-06-29
SOUTHWEST UNIV
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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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  • A method for molecular cloning and identification of silkworm genes
  • A method for molecular cloning and identification of silkworm genes
  • A method for molecular cloning and identification of silkworm genes

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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] Such as 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 tran...

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Abstract

The invention belongs to the field of biology and discloses a method for molecular cloning and identification of a silkworm gene. The sequence of the silkworm gene is SEQ ID NO: 1; it is named BmDHODH. The present invention uses leflunomide to treat silkworm cells, which has a great inhibitory effect on BmDHODH expression and cell proliferation; as a component in pyrimidine synthesis, BmDHODH is an essential biological enzyme closely related to cell growth and proliferation, and it may become The new gene target for killing Lepidoptera pests provides new insights into eco-friendly pest control, and provides new targets and theoretical methods for the development of environmentally friendly insecticides; The model, BmDHODH of the silkworm, can enrich the basic knowledge of Lepidoptera pests and provide new methods for environmental 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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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12N15/10C12Q1/6888
CPCC12N9/001C12N15/10C12Q1/6851C12Y103/01014C12Q2531/113C12Q2561/113
Inventor 崔红娟赵二虎蒋小兰张奎杨丽群
Owner SOUTHWEST UNIV
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