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Application of G6PDH gene segment taken as stable expression reference gene of Bactrocera minax under intimidation condition

A technology of Bactrocera citrus, BM-G6PDH-F1, applied in the field of molecular biology, can solve problems such as errors

Active Publication Date: 2013-10-16
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, blindly using a housekeeping gene as an internal reference may lead to wrong or even opposite conclusions

Method used

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  • Application of G6PDH gene segment taken as stable expression reference gene of Bactrocera minax under intimidation condition
  • Application of G6PDH gene segment taken as stable expression reference gene of Bactrocera minax under intimidation condition
  • Application of G6PDH gene segment taken as stable expression reference gene of Bactrocera minax under intimidation condition

Examples

Experimental program
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Embodiment 1

[0017] Example 1: Expression analysis of the internal reference gene BM-G6PDH of Bactrocera citrus under different stress conditions

[0018] 1. Extraction of total RNA from Bactrocera citrus under different stress conditions:

[0019] Take the larvae of Bactrocera citrus and its different stress times (12h, 36h, 48h, 60h) at 4°C, and the adults and one of them after different stress times (1h, 3h, 5h) at 35°C and irradiation (90Gy). Put it into a 1.5mL centrifuge tube, add enough liquid nitrogen and use a grinding rod to grind it into powder; add 1mL Trizol to the centrifuge tube that has been ground into powder, shake and mix well, and place it at room temperature for 5min; after standing still, add 0.2mL Trizol Chloroform, fully oscillate 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, fully shake and mix for 15 seconds, place at room tempera...

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Abstract

The invention relates to the field of molecular biology, particularly to the application of a G6PDH gene segment taken as a stable expression reference gene of a Bactrocera minax under the intimidation condition. According to the invention, the selected G6PDH ( glucose-6-phosphate dehydrogenase) reference gene is subjected to semiquantitative RT-PCR technology detection, as a result, the fact that both an imago under high temperature of 35 DEG C and irradiation condition and a larva under low temperature of 4 DEG C intimidation condition can express stably is shown, so that the application lays a foundation for the research of Bactrocera minax function and the expression of Bactrocera minax under different intimidation conditions.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to the application of the G6PDH gene fragment as an internal reference gene stably expressed under the stress condition of Bacteropherus citrus. Background technique [0002] Insects are ectotherms and are very sensitive to temperature. Under unfavorable high and low temperature stress conditions, insects often prefer to improve their own physiological adaptability as their preferred survival strategy. Therefore, studying the adaptation mechanism of insects to high and low temperatures can provide a deeper understanding of the survival selection strategies adopted by species in stressful environments. Organisms perceive external temperature stimuli, transmit signals to the body, and present a series of gene stress expressions in the body to cope with external stress. In order to better understand the mechanism of insect adaptation to high and low temperature at the gene level, it ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
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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