Specific probe for detecting hemocyanin gene expression of eriocheir sinensis and application of specific probe

Abstract

The invention discloses a specific probe for detecting hemocyanin gene expression of Eriocheir sinensis and application of the specific probe. Based on the principle that a probe sequence is specifically combined with mRNA of a target gene in proportion, a fluorescence in-situ hybridization technology is combined with image flow cytometry, different groups are distinguished according to whether fluorescence signals exist in cells or not, functional differences are analyzed according to signal strength, and the target gene can be identified according to the functional differences. And whether different genes are expressed in the same cell or not can be known by using multicolor fluorescence co-localization. Cell classification serves for diagnosis, function research and other requirements. Conventional methods mostly adopt antibody recognition to classify cells, but the current situation that aquatic animals lack commercially available antibodies limits the application. The method does not need an antibody, starts from a gene sequence, prepares a fluorescently-labeled short probe, is mild in hybridization condition, can carry out quantitative analysis at a complete single cell level, and has a good application prospect in cell classification and other aspects.

Description

[0001] This patent application was completed by the School of Life Sciences of Tianjin Normal University / Tianjin Key Laboratory of Animal and Plant Resistance, and was awarded the Tianjin Application Basic and Frontier Technology Research Program (19JCYBJC29700) and the Tianjin Key Laboratory of Aquatic Ecology and Aquaculture Open Fund (TJAE2015005). . technical field [0002] The invention belongs to the field of detection technology, and relates to a single-cell-level high-throughput gene based on the proportional specific combination of a probe sequence and a target gene target sequence, combining fluorescence in situ hybridization technology and flow cytometry detection technology Novel automated assays for expression measurement and cell classification. This method does not need to use antibodies to classify mitten crab blood cells, and only needs to obtain partial sequence information of the target gene, and then the probe sequence can be synthesized. The shorter probe ...

AI Technical Summary

Problems solved by technology

For a long time, the research on immune cells of aquatic animals has mainly relied on manual methods such as staining and microscopic examination. The technical bottleneck of high-throughput automated classification and analysis has also greatly affected the development of related research.

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