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Method for obtaining transgenic plant for producing degradable plastics

A technology of transgenic plants and degrading plastics, applied in the fields of botanical equipment and methods, plant products, angiosperms/flowering plants, etc., can solve the problem that PHB specific expression cannot be achieved, the expression level is low, and the toxicity of PHB to plants cannot be effectively avoided. role and other issues to achieve the effect of solving the energy crisis

Inactive Publication Date: 2010-05-19
山西九龙湾农林科技开发有限公司
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Problems solved by technology

At present, the promoters used to produce PHB mainly include soybean β-phaseolin promoter, soybean glutenin Gtl promoter, rapeseed napinB protein promoter, rapeseed NapA gene promoter, etc., but these promoters have low expression levels and gene silence. , and cannot realize the specific expression of PHB in seeds, and cannot effectively avoid the toxic effect of PHB on plants

Method used

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  • Method for obtaining transgenic plant for producing degradable plastics
  • Method for obtaining transgenic plant for producing degradable plastics
  • Method for obtaining transgenic plant for producing degradable plastics

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Embodiment Construction

[0019] 1. Cloning of the faba bean Leg gene promoter and construction of its vector:

[0020] (1) Design degenerate primers according to the faba bean genome sequence data, and use the faba bean total DNA as a template to achieve amplification by PCR. The amplification primers are:

[0021] Primer P1:

[0022] 5 `cttgctgtg cagctcgcct agtgaagct 3`

[0023] Primer P2:

[0024] 5`acccagata caatcgctgt taacgtagt 3`

[0025] 94°C for 5min; 35 cycles of 94°C for 30s, 55°C for 1min, and 72°C for 2min; 72°C for 10min. A DNA fragment of about 3.2 kb can be generated, and the sequenced result is shown in the sequence list.

[0026] On this basis, the following two primers were designed and synthesized to realize the integration of HindIII and Sal I restriction sites:

[0027] Primer L1:

[0028] 5'-GCG aagctt gtgtcgctcagctcgcctagtg-3'

[0029] Hind III

[0030] Primer L2:

[0031] 5'-cgc gtcga agcgacaattgcaggcagca-3'

[0032] Sal I

[0033] A HindIII site and a Sal I site...

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Abstract

The invention relates to the technical field of genetic engineering, in particular to a method for obtaining a transgenic plant for producing degradable plastics, which solves the problems in damaging a plant material by polyester, overcoming gene silencing, improving PHB expression, and the like. The method comprises the following steps: (1) cloning a promoter and constructing a promoter vector; (2) connecting the promoter vector to phbA, phbB and phbC genes so as to construct an expression vector; and (3) introducing the expression vector into a plant receptor to obtain the transgenic plant, wherein the promoter is seed-specificity promoter, is an Leg gene promoter cloned by using the total DNA of horse bean as a template, and has a nucleotide sequence as shown by a sequence table, the constructed promoter vector is Leg-GUS, the constructed expression vector is pSCAB, and the plant receptor is xanthium sibiricum. The PHB average accumulation in a cultured xanthium sibiricum seed exceeds 28 percent of the green weight, which provides a practical method and experiences for researches on producing the degradable plastics by using the transgenic plant, and initiates a new situation for the researches on producing the degradable plastics by using the transgenic plant in China.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for obtaining transgenic plants for producing degradable plastics. Background technique [0002] With the development of industry, plastics, steel, cement, and wood have become new basic material industries that go hand in hand, and their application fields far exceed the other three industries. In recent years, China's demand for plastic packaging materials has increased significantly, and has penetrated into various sectors of the national economy and all areas of people's lives. At present, the world's annual consumption of plastics reaches 240 million tons, with an annual growth rate of about 4%; China's annual plastic consumption has reached more than 40 million tons, and it is expected to continue to grow at an average annual rate of at least 8% in the next few years. However, most of disposable plastic packaging materials, disposable daily necessities...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/10
Inventor 马小军赵峰
Owner 山西九龙湾农林科技开发有限公司
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