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BM-EF1-alpha segment stably expressed in different development stages of bactrocera minax and application thereof

A citrus fruit fly, stable expression technology, applied to the field of the internal reference gene BM-EF1-α fragment, can solve the problem of easy changes in the expression of the internal reference gene

Active Publication Date: 2013-09-04
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in some studies, it was found that not all commonly used internal reference genes can be stably expressed under any circumstances, and the expression of some internal reference genes is prone to change under specific conditions

Method used

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  • BM-EF1-alpha segment stably expressed in different development stages of bactrocera minax and application thereof
  • BM-EF1-alpha segment stably expressed in different development stages of bactrocera minax and application thereof
  • BM-EF1-alpha segment stably expressed in different development stages of bactrocera minax and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Analysis of the expression of the reference gene BM-EF1-α in Bactrocera citrus at different developmental stages

[0021] 1. Extraction of total RNA from Bactrocera citrus at different developmental stages:

[0022] Take 30 eggs, larvae / pupa (1 day, 90 days, 160 days) / adults (male and female) of Bactrocera citrus and put them into a 1.5mL centrifuge tube, add enough liquid nitrogen and use a grinding rod to grind them into Powder; add 1mL Trizol to the powdered centrifuge tube, shake and mix well, and place at room temperature for 5 minutes; after standing still, add 0.2mL of chloroform, fully shake and mix for 15 seconds, place at room temperature for 5 minutes, and centrifuge at 12,000 rpm for 15 minutes at 4°C; Draw the supernatant into another EP tube, add 0.2mL chloroform, shake and mix well for 15s, place at room temperature for 5min, and centrifuge at 12000 rpm for 15min at 4°C; take the supernatant into another EP tube and add an equal volume of iso ...

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Abstract

The invention relates to the field of molecular biology, and in particular relates to an endogenous reference gene BM-EF1-alpha segment of bactrocera minax and an application thereof. The nucleotide sequence of the gene segment is shown by SEQ ID No.1. According to the BM-EF1-alpha of bactrocera minax, provided by the invention, the nucleotide sequence with 224bp can be used as a functional gene for studying bactrocera minax in future and an endogenous reference gene for gene expression in different development stages.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to an internal reference gene BM-EF1-α fragment stably expressed in different development stages of Bactrocera citrus and its application. Background technique [0002] Endogenous references gene (Endogenous references gene) is an internal reference, which refers to a gene that is constantly expressed under a certain condition, and serves as a reference for detecting the expression level change of the target gene under a specific condition. Many commonly used internal reference genes are expressed in various cells of organisms, and they are of great significance in maintaining and regulating the basic life activities of cells. However, in some studies, it was found that not all commonly used internal reference genes can be stably expressed under any circumstances, and the expression of some internal reference genes is easy to change under certain conditions. Therefore, when choosin...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/12
Inventor 吕志创王刘豪张桂芬周忠实万方浩郭建英
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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