Endogenous reference gene BM-alpha-TUB stably expressed in different development stages of bactrocera minax and application thereof

A stable expression technology of Bactrocera citrus, which is applied to the internal reference gene BM-α-TUB and its application fields, and can solve the problems of expression changes

Active Publication Date: 2015-03-11
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the expression of some internal reference genes may change under certain conditions

Method used

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  • Endogenous reference gene BM-alpha-TUB stably expressed in different development stages of bactrocera minax and application thereof
  • Endogenous reference gene BM-alpha-TUB stably expressed in different development stages of bactrocera minax and application thereof
  • Endogenous reference gene BM-alpha-TUB stably expressed in different development stages of bactrocera minax and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Expression analysis of the internal reference gene BM-α-TUB of 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, oscillate fully, and place at room temperature for 5min; after standing still, add 0.2mL of chloroform, fully oscillate and mix for 15s, place at room temperature for 5min, and centrifuge at 12,000 rpm for 15min at 4°C ;Pipe 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 Isopropanol, shake and ...

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Abstract

The invention relates to the field of molecular biology, and in particular relates to an endogenous reference gene BM-alpha-TUB stably expressed in different development stages 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-alpha-TUB of bactrocera minax provided by the invention, the nucleotide sequence with 178bp 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-α-TUB stably expressed in different development stages of Bactrocera citrus and its application. Background technique [0002] Citrus Bactrocera (Bactrocera (Tetradacus) Minax) belongs to Arthropoda, Insecta, Diptera, Tephritidae, and is commonly called as citrus maggot. It is one of the important pests that harm citrus fruit trees. It is also an international phytosanitary pest. Now it is widely distributed in citrus producing areas such as Southwest China, South China, Central South and East China. [0003] To complete a life cycle, Bacterophagus citrus needs to go through four stages: egg, larva, pupa and adult. It is a holometabolous insect with one generation per year. The overwintering generation adults begin to emerge in late April each year. After the adults mate, the female adults Lay eggs in young fruit successively from early June to mid...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/12
Inventor 吕志创王刘豪周忠实张桂芬万方浩郭建英
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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