Ceratopteris pteridoides gene for resisting human immunologic deficiency disease toxoprotein congeners, and preparation method and application thereof

A technology for human immunodeficiency and homologues, applied in botany equipment and methods, biochemical equipment and methods, antiviral agents, etc., can solve problems such as expensive patent fees and limited application development research

Inactive Publication Date: 2010-09-22
WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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Problems solved by technology

The CVNH gene without independent intellectual property rights will force my country's biomedical industry to pay expensive patent fees in the future commercial use, and will greatly limit related application development research

Method used

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  • Ceratopteris pteridoides gene for resisting human immunologic deficiency disease toxoprotein congeners, and preparation method and application thereof
  • Ceratopteris pteridoides gene for resisting human immunologic deficiency disease toxoprotein congeners, and preparation method and application thereof
  • Ceratopteris pteridoides gene for resisting human immunologic deficiency disease toxoprotein congeners, and preparation method and application thereof

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Embodiment Construction

[0033] A method for preparing a water fern human immunodeficiency virus protein homologue gene, comprising the steps of:

[0034] 1. Total DNA extraction:

[0035] Cut the young leaves of the water fern sporophyte, put them into liquid nitrogen immediately, use the improved CTAB method to extract the total DNA, and use a UV spectrophotometer and agarose gel electrophoresis with a mass ratio of 1.0% to detect the concentration and purity of the extracted DNA , stored at -20°C or -80°C.

[0036] 2. Amplification of the middle fragment of CVNH gene:

[0037] Using the extracted total DNA as a template, use a pair of specific primers for PCR amplification. The primer sequences are:

[0038] CVNH-F: 5'-GTGGGCGTCTAGCGATTTCCTTT-3'

[0039] CVNH-R: 5'-ATCATCCGCTGCTTGCTTCTTCG-3'

[0040] PCR reaction system: 20 μl, 40 pmol each primer, 1 pmol each dNTP, 2.0 μl 10×PCR reaction buffer, 0.2 μl Taq enzyme (5U μl -1 ). The reaction conditions were pre-denaturation at 94°C for 5 min; f...

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Abstract

The invention discloses a ceratopteris pteridoides gene for resisting human immunologic deficiency disease toxoprotein congeners, and a preparation method and application thereof. The preparation method comprises the following steps: 1, extracting total DNA by using an improved CTAB method, and preserving; 2, amplifying the intermediate segment of the ceratopteris pteridoides gene for resisting human immunologic deficiency disease toxoprotein congeners; 3, amplifying 5' flanking sequence of the ceratopteris pteridoides gene for resisting human immunologic deficiency disease toxoprotein congeners; 4, amplifying 3' flanking sequence of the ceratopteris pteridoides gene for resisting human immunologic deficiency disease toxoprotein congeners; and 5, recycling and cloning the PCR product: recycling the intermediate segment of the target gene, and 5' and3' flanking sequence segments, transforming, culturing and selecting white colonies; and 6, judging the clone, measuring the sequence and splicing. The gene can be used for regulation and control on expression of potential antivirus genes in plant research, carrying out research into plant antivirus gene engineering, and further establishes a ceratopteris pteridoides CVNH protein preparation system, thereby finally realizing large-scale industrial production.

Description

technical field [0001] The present invention relates to the field of biotechnology. Specifically, the present invention relates to a gene of water fern anti-human immunodeficiency virus protein homologue, a preparation method of a human immunodeficiency virus protein homologue gene, and the protein encoded by the gene in anti-human immunodeficiency virus Application in immunodeficiency virus medicine. Background technique [0002] AIDS has become an infectious disease that seriously threatens the global social and economic security, and the situation in our country is not optimistic. According to the Ministry of Health, UNAIDS and the World Health Organization at the 2008 World AIDS Day press conference on the AIDS epidemic in China, as of September 30, my country had reported a total of 264,302 AIDS cases, including 77,753 patients and 34,864 deaths. At present, chemotherapy is mostly used in the treatment of AIDS, which is expensive and prevents many ordinary patients fr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/10C07K14/415A61K38/16A61P31/18
Inventor 齐小琼苏应娟王艇杨永霞
Owner WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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