Method for large-scale quick separation of insect peritrophic membranes

A peritrophic film and mass-volume technology, applied to animal cells, etc., can solve problems such as heavy workload

Inactive Publication Date: 2013-04-24
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

However, the peritrophic membrane of insects is very thin, and it takes a lot of skill and patience to dissect and separate the peritrophic membrane of an insect from the midgut. There is still food in the isolated peritrophic membrane tissue. In the case of ensuring that the peritrophic membrane protein is not degraded, it is another technical difficulty to quickly clean the food inside, and the fifth instar larvae, which is the size of a cotton bollworm, only has a dry weight of 53.45 μg after separation of the peritrophic membrane. quite big

Method used

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Examples

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Embodiment Construction

[0016] In order to clearly understand the present invention, the present invention will be further described in detail below in conjunction with the examples.

[0017] This embodiment takes the rapid separation of large batches of peritrophic membranes of cotton bollworm as an example. It should be noted that the present invention is not limited to this embodiment. In the following examples, the preparation of cotton boll larvae specimens was obtained by selecting healthy 5th instar cotton boll larvae and rapidly freezing (15 minutes or more) at 0°C.

[0018] The specific implementation steps are as follows:

[0019] 1. Take 100-200 specimens of 5th instar cotton boll larvae and put them in an ice bath to cool down;

[0020] 2. Take 2 mL of EP tubes corresponding to the number of 5th instar cotton boll larvae specimens, add 2 mL of 0.7% NaCI physiological saline that has been cooled in advance at 0°C to each EP tube, and put these EP tubes filled with physiological saline Al...

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Abstract

The invention discloses a method for large-scale quick separation of insect peritrophic membranes. The method can separate peritrophic membrane organizations from insect midgut in a short time, and then food residues therein are quickly cleaned, so that a large amount of purified peritrophic membrane organizations are obtained; the efficiency is improved by 16 times when compared with that of the general method; and the convenience is provided to the subsequent research of proteins in the peritrophic membranes.

Description

technical field [0001] The invention relates to a technique for separating insect tissues, in particular to a new method for rapidly separating a large number of insect peritrophic membranes. Background technique [0002] The peritrophic membrane (PM) is a thick and uniform acellular structure film secreted by insect midgut cells, extending from the front of the midgut to the hindgut. The peritrophic membrane is mainly composed of protein and chitin. It has many functions such as protecting midgut epithelial cells and preventing the invasion of pathogens. It is the first natural protective barrier for insects to resist the invasion of pathogenic microorganisms ingested with food. Potential target sites for pest control. Therefore, in recent years, studies on the assembly mechanism of chitin and protein in the peritrophic membrane, the molecular mechanism of the interaction between pathogenic microorganisms and the peritrophic membrane, and the prevention of insect resistan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/07
Inventor 梁革梅李怡萍郭予元魏纪珍
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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