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Gobiocypris rarus cell line, and culture method and induced transformation method thereof

A kind of technology of rare crucian carp and culture method, applied in the field of cell biology, can solve problems such as unseen application, and achieve the effect of rapid cell growth and stable shape

Pending Publication Date: 2021-03-26
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the rare gobi carp is only used in the detection of aquatic ecotoxicity, and there is no report on its application in the study of aquatic animal cell transformation.

Method used

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  • Gobiocypris rarus cell line, and culture method and induced transformation method thereof
  • Gobiocypris rarus cell line, and culture method and induced transformation method thereof
  • Gobiocypris rarus cell line, and culture method and induced transformation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0080] Example 1 of the present invention discloses a rare minnow carp swim bladder cell line, the preservation number is CCTCC NO: C2019243.

[0081] Specifically, its classification is named as the rare goblin carp swim bladder cell line GrSB, which was preserved in the China Type Culture Collection Center on September 26, 2019, and proved to be alive. The preservation registration number is CCTCC NO: C2019243, and the preservation address is Wuhan, China .Wuhan University.

[0082] Embodiment 1 of the present invention also provides a method for cultivating the rare gobi carp swim bladder cell line as described above, which specifically includes the following steps:

[0083] (1) Primary culture of rare squid carp swim bladder cells

[0084] (11) Soak the fresh and healthy rare squid in 75% alcohol for 1-2 minutes, then place it in an ultra-clean bench to aseptically remove the swim bladder tissue and remove the components of the surface membrane of the swim bladder tissue,...

Embodiment 2～5

[0121] Embodiments 2 to 5 of the present invention also provide a method for inducing transformation of the rare gobi carp swim bladder cell line as disclosed in Embodiment 1, which specifically includes the following steps:

[0122] (A) Preparation of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) solution

[0123] First, dissolve N-methyl-N'-nitro-N-nitrosoguanidine in deionized water to prepare a 10mg / mL mother solution, and then prepare N-methyl- N'-nitro-N-nitrosoguanidine cell culture solution, the specific concentration is shown in Table 1 below;

[0124] (B) N-methyl-N'-nitro-N-nitrosoguanidine induces swim bladder cells of rare gobi carp

[0125] The rare gobi carp swim bladder cells subcultured to the 5th generation in Example 1 were cultured to the exponential growth phase, and then N-methyl-N'-nitro-N-nitrosoguanidine cell culture solution was added at 27°C Cultivate for 24-48 hours to obtain induced cultured cells;

[0126] (C) Low serum culture

[0127] Take the...

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Abstract

The invention discloses a gobiocypris rarus cell line with the preservation number of CCTCC NO: C 2019243, and discloses a construction and culture method of the gobiocypris rarus cell line. The construction and culture method specifically comprises the steps of primary culture of gobiocypris rarus cells, subculture of the gobiocypris rarus cells and cryopreservation and recovery of the cells. Theinvention also discloses an induced transformation method of the gobiocypris rarus cell line. The induced transformation method specifically comprises the following steps of preparing an N-methyl-N'-nitro-N-nitrosoguanidine solution, inducing the gobiocypris rarus cells by N-methyl-N'-nitro-N-nitrosoguanidine, and performing low-serum culture after induction. The gobiocypris rarus cell line, andthe culture method and the induced transformation method of the gobiocypris rarus cell line disclosed by the invention provide the basis for the application of gobiocypris rarus in an aspect of aquatic animal transformation research, and the application of the gobiocypris rarus to biological experiments can be expanded.

Description

technical field [0001] The invention relates to the technical field of cell biology, and more specifically relates to a rare goblin carp swim bladder cell line and a culture method and induction transformation method thereof. Background technique [0002] The rare gobiocypris rarus (Gobiocypris rarus) belongs to the genus Gobiocypris, which is endemic to China and is mainly distributed in the upper reaches of the Yangtze River in Sichuan Province. Rare gobi crucian carp has a small body size (adult fish 30mm-80mm), can adapt to growth in a wide temperature range (0-30°C), sexual maturity and short life cycle, long breeding season, high spawning frequency, and high spawning volume. Large size, transparent eggs, easy to observe, and sensitivity to environmental pollutants. These characteristics make the rare gobi crucian carp a good test organism for aquatic ecotoxicology, and is recommended as a test organism in China for use in aquatic ecotoxicity testing. [0003] However,...

Claims

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Application Information

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IPC IPC(8): C12N5/071A01N1/02C12N15/01C12R1/91
CPCA01N1/02C12N5/0602C12N15/01
Inventor 王英英王庆尹纪元李莹莹吴斯宇石存斌任燕
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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