Method for cloning rice auxin induced protein gene

A technology for inducing protein and auxin, applied in the fields of biochemical equipment and methods, genetic engineering, plant genetic improvement, etc., can solve the problems that restrict the high-level development of agricultural biotechnology breeding, lack and other problems, and achieve easy implementation steps and implementation conditions. The effect of loose, simple implementation steps

Inactive Publication Date: 2009-07-29
TIANJIN AGRICULTURE COLLEGE
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Problems solved by technology

However, the serious lack of important functional genes with independent intellectual property rights is a "bott

Method used

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Embodiment Construction

[0022] Below in conjunction with the preferred embodiment, the specific implementation steps provided according to the present invention are described in detail as follows:

[0023] (1) Preparation of rice transformed receptors

[0024] Dehull the mature seeds of rice variety Nipponbare, first soak in 70% alcohol for 1 minute, then sterilize with 0.1% mercuric chloride solution for 20 minutes, then rinse with sterile water for 3 times, blot dry and inoculate into NB D Callus induction was carried out on the culture medium at a culture temperature of 25±1° C. After 1 week, the callus was picked for subculture, and after 2 weeks of subculture, the callus was taken out for transformation.

[0025] (2) Genetic transformation of rice

[0026]Pick a single colony of Agrobacterium into YEP liquid medium containing 50mg / L kanamycin and 20mg / L rifampicin, culture with shaking at 28°C for 14 hours, so that the concentration of the bacteria solution reaches the OD value of 0.5, and then...

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Abstract

The invention relates to a method for using rice auxin to induce protein genes to be cloned, which is characterized in that the implementation steps comprise: (1) the preparation of a rice transformation receptor; (2) the genetic transformation of the rice; (3) the screening of kanamycin-resistant callus tissue and the regeneration of plants; (4) the screening of the mutant of a T-DNA inserted progenies; (5) Tail-PCR; (6) the comparison and analysis of sequences on the Internet. In the invention, a rice mutant with a short plant height is obtained when carrying out rice functional genome research by using T-DNA label method; the lateral neighboring sequences of the mutant are researched by using TAIL-PCR technology; meanwhile, the position where the mutant T-DNA inserts the rice genome is arranged on the No. 4 chromosome of the rice by the comparison on the databases of NCBI and TIGR on the Internet; moreover, the rice BAC clone (OSJNBa0084K01) of the position is found out. The T-DNA is inserted between the two genes of the clone by analyzing the clone. Known functional genes with a very high homology with BAC cloning code amino acid sequence are forecasted by the sequence comparison on the Internet.

Description

technical field [0001] The invention relates to a method for applying rice functional genomics, in particular to a method for cloning a rice auxin-induced protein gene. Background technique [0002] Rice is one of the three major food crops. More than one-third of the world's population uses it as the main source of food. It plays a pivotal role in the national economy. With the continuous increase of the world population and the increase of the demand for food, the annual demand for rice is constantly increasing. Today, with the shrinking land area, how to make rice production stable and high to alleviate the impact of the increase in population on rice production The pressure brought by it is a difficult problem faced by the world, and it is also a major scientific research topic in front of scientific researchers. Although traditional genetic research and conventional breeding have made important contributions to the improvement of rice yield and quality, due to the limi...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12N15/29
Inventor 裴忠有孙守钧李子芳罗峰
Owner TIANJIN AGRICULTURE COLLEGE
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