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shRNA for inhibiting expression of phyllotreta striolata arginine kinase protein as well as preparation method, primer and application of shRNA

A technology of arginine kinase protein and flea beetle, applied in the field of genetic engineering, can solve problems such as ecological environmental pollution, increased resistance of pests, and impact on food safety

Pending Publication Date: 2020-09-11
SHENZHEN UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no effective control method for flea beetle. Spraying chemical pesticides is still the main control method in production. However, the long-term and large-scale use of chemical pesticides not only causes ecological environment pollution, affects food safety, but also leads to pest resistance. Gradually increase the potency

Method used

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  • shRNA for inhibiting expression of phyllotreta striolata arginine kinase protein as well as preparation method, primer and application of shRNA
  • shRNA for inhibiting expression of phyllotreta striolata arginine kinase protein as well as preparation method, primer and application of shRNA
  • shRNA for inhibiting expression of phyllotreta striolata arginine kinase protein as well as preparation method, primer and application of shRNA

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preparation example Construction

[0040] The embodiment of the present invention provides a method for preparing an interfering shRNA that inhibits the expression of the arginine kinase protein of the yellow-cured strips, which includes the steps of:

[0041] According to the sequence of the target gene, design the interfering shRNA sequence;

[0042] According to the interfering shRNA sequence, design a pair of primer sequences;

[0043] Use the designed pair of primers to generate and prepare shRNA transcription templates through filling reactions;

[0044] The prepared shRNA transcription template is used to prepare the interfering shRNA through in vitro transcription.

[0045] In one embodiment, the filling reaction system includes: a designed pair of primers, dNTPs, DNA polymerase and the like.

[0046]In one embodiment, the in vitro transcription system includes: shRNA transcription template, T7 RNA polymerase buffer, NTPs, T7 RNA polymerase and the like.

[0047] The embodiment of the present inventi...

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Abstract

The invention discloses shRNA (short hairpin Ribonucleic Acid) for inhibiting expression of phyllotreta striolata arginine kinase protein as well as a preparation method, a primer and application of the shRNA. The interfering shRNA for inhibiting the expression of the phyllotreta striolata arginine kinase protein sequentially comprises a target sequence, an annular structure sequence and a complementary sequence of the target sequence, wherein the target sequence is GGAGUUCGAUGCCGUUAAGGA. The interfering shRNA for inhibiting the expression of the phyllotreta striolata arginine kinase protein provided by the invention can stably, efficiently, remarkably and specifically inhibit the expression of mRNA (messenger ribonucleic acid) of a coding light capture protein gene in mikania micrantha.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, and in particular to an interfering shRNA that specifically targets and inhibits the expression of arginine kinase protein in japonica japonica, a preparation method, primers and applications. Background technique [0002] Flea beetle is widespread worldwide and is the number one vegetable pest in Guangdong. At present, there is no effective control method for flea beetle. Spraying chemical pesticides is still the main control method in production. However, the long-term and large-scale use of chemical pesticides not only causes ecological environment pollution, affects food safety, but also leads to pest resistance. The medicinal properties are gradually enhanced. Therefore, it is very urgent to develop green prevention and control technology. [0003] RNA interference (RNAi) refers to the phenomenon of efficient and specific degradation of homologous mRNA induced by double-strande...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/10C12N15/11A01N57/16A01P7/04
CPCC12N15/1137A01N57/16C12N2310/14C12N2310/531
Inventor 陈伟钊莫亦琳钟路遥王晓彦莫蓓莘
Owner SHENZHEN UNIV
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