Clonal strain of cabbage looper cell line and application thereof

A technology of trichoplusiasis and cell lines, applied in the field of insect cell line clones, can solve the problems of cell agglomeration, unsuitable virus and recombinant protein production, etc., and achieve the effect of reducing production costs

Inactive Publication Date: 2012-03-07
QINGDAO AGRI UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is that the currently used cell lines Sf-21, Sf-9 and HighFive are not suitable for suspension mass culture due to their strong attachment to the wall during the culture process, thus affecting the production of viruses and recombinant proteins. Clone strain of Trichoplusia spp. cell line, thereby overcoming the problem of cell agglomeration in suspension culture and further increasing the expression level

Method used

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  • Clonal strain of cabbage looper cell line and application thereof
  • Clonal strain of cabbage looper cell line and application thereof
  • Clonal strain of cabbage looper cell line and application thereof

Examples

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Embodiment 1

[0023] Example 1 Isolation and identification of cell clone QB-TN9-4S-CL-B (QB-CL-B)

[0024]1.1 Establishment of Trichoplusia cell line QB-TN9-4S: according to the method of Ming-Juan Meng (2008, Insect Science, 15: 423-428), fresh eggs of Trichoplusia populations cultured in the laboratory were obtained, respectively After surface disinfection with 10% sodium hypochlorite and 75% alcohol for 5 minutes and washing twice with TNM-FH (GIBCO, USA) medium, eggs were transferred to a cell strainer (BD Falcon, USA) and the strainer Put it into a plate with 10ml medium, grind the cells fully, dilute the egg tissue filtrate to different concentrations, and distribute it in 25cm 2 culture flask (Corning, New York, USA). Incubate at 28°C. Half of the medium was replaced with fresh medium every 2 weeks. After 5 months, when the formed cells covered the bottom of the bottle, the first subculture was carried out. After the cells grew stably, they were subcultured twice a week.

[0025...

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Abstract

The invention discloses a clonal strain of a cabbage looper cell line and application thereof. The microorganism preservation number of the clonal cell strain is CCTCC C201046. The clonal strain cannot attach the wall tightly so as not to clump in the suspension culture, which is beneficial to the attachment and the infection of viruses and the significant improvement of the yield of viruses and recombinant proteins. Compared with the wild-type cells, the clonal strain of the invention has high expression level, and the recombinant protein expression of the clonal strain is 1.2-3.0 times that of the wild-type cells. The clonal strain can be applicable for cultivating heavy (QB-TN9-4s) and High-Five cell histone having a wide international application at present or insecticidal viruses. The clonal strain is as high as the High-Five cells in the virus sensitivity and the virus yield. However, the polyhedron yield of the clonal strain is obviously higher than that of another widely used SF-9 cells and is 2.9 times that of the SF-9 cells.

Description

technical field [0001] The present invention relates to a clone of an insect cell line, especially a new clone of a Trichoplusia spp. cell line. The present invention also relates to the use of the clone in preparing an insecticidal virus or recombinant protein, belonging to insect cell biology and biology. technology field. Background technique [0002] Since Grace established the first insect cell line in 1962 (Grace T.D.C.1962, Nature, 195: 788-789), 600 insect cell culture medium or improved Grace medium have been used to establish 600 insect cell lines in more than 100 species. Yu strains of insect cell lines (Chih-YuWu, 2006, J. Invert. Pathol., 93: 186-191). Insect cell lines were originally used in the research of insect viruses. With the continuous development of research, the scope of application of insect cells has been continuously expanded, especially the invention of baculoviruses-insect cell expression system (Thebaculoviruses-insect cell expression system), ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/07C12N7/00C12N9/38C12N9/16C12N15/63C07K2/00C12R1/91
Inventor 李国勋李天龙姜磊马明李淑文吴艳蕾
Owner QINGDAO AGRI UNIV
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