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cRNA (complementary ribonucleic acid) in-situ hybridization probe of glutamate transporter-1 gene and preparation method of cRNA in-situ hybridization probe

A technology of transporter and in situ hybridization, which is applied in the field of biomedicine, can solve the problems that are still blank, achieve the effect of clear specificity and display effect

Inactive Publication Date: 2013-12-25
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

In terms of morphological research, the detection of the mRNA level of the glutamate transporter-1 gene in living animals shows that it is still blank

Method used

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  • cRNA (complementary ribonucleic acid) in-situ hybridization probe of glutamate transporter-1 gene and preparation method of cRNA in-situ hybridization probe
  • cRNA (complementary ribonucleic acid) in-situ hybridization probe of glutamate transporter-1 gene and preparation method of cRNA in-situ hybridization probe

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Embodiment Construction

[0048] The present invention is described in further detail below in conjunction with accompanying drawing:

[0049] see figure 1 with figure 2 , the preparation method of glutamate transporter-1 gene cRNA in situ hybridization probe, according to the following steps:

[0050] (1) According to the gene bank sequence of glutamate transporter-1, we designed primers specific to glutamate transporter-1, and this pair of primers amplified a 280bp fragment of glutamate transporter-1 as glutamate Translocator-1 specific probe sequences;

[0051] (2) Construct the probe plasmid of the cRNA in situ hybridization of glutamate transporter-1; After the probe plasmid is transformed into bacteria, the bacteria are cultured and then sequenced;

[0052] (3) preparing the cRNA in situ hybridization probe of glutamate transporter-1;

[0053] (4) In situ hybridization of cRNA probe for detecting glutamate transporter-1.

[0054] Probe design:

[0055] According to the gene bank sequence o...

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Abstract

The invention discloses a cRNA (complementary Ribonucleic Acid) in-situ hybridization probe of a glutamate transporter-1 gene and a preparation method of the cRNA in-situ hybridization probe. The preparation method comprises the following steps of: (1) designing a glutamate transporter-1 specific primer according to a Genebank sequence of the glutamate transporter-1 and amplifying a 280 bp glutamate transporter-1 fragment for the primer to serve as a glutamate transporter-1 specific probe sequence; (2) constructing cRNA in-situ hybridization probe plasmid of the glutamate transporter-1, converting the probe plasmid into bacteria, culturing the bacteria and then sequencing; (3) preparing the cRNA in-situ hybridization probe of the glutamate transporter-1; and (4) detecting in-situ hybridization of the cRNA probe of the glutamate transporter-1.

Description

Technical field: [0001] The invention belongs to the field of biomedicine and relates to a probe and a preparation method thereof, in particular to a cRNA in situ hybridization probe of glutamate transporter-1 gene and a preparation method thereof. Background technique: [0002] Glutamate is the most important neurotransmitter in the central nervous system of mammals. It mainly binds to two families of glutamate receptors, ionophilic and metabotropic, to activate them. If the concentration of glutamate in the extracellular area is too high, excessive excitation of glutamate receptors can lead to nerve cell death. Since there is no enzyme that can decompose glutamate outside the cell, the released glutamate is almost entirely dependent on neurons and glia The uptake of glutamate transporter (EAAT) on the cell membrane keeps the concentration relatively stable, and EAAT also provides a source of glutamate for many metabolic pathways in the body. Glutamate transporters are div...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11C12N15/10
Inventor 魏燕燕黄静陈晶王文李云庆武胜昔
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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