Plant expression vector for the specific expression of epatitis B surface antigen in fruits

A plant expression vector and hepatitis B virus technology, which can be used in the introduction of foreign genetic material using vectors, recombinant DNA technology, etc., can solve the problems of inhibiting plant growth, affecting the growth process of plant cells, and low expression levels.

Inactive Publication Date: 2010-02-10
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI +2
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Problems solved by technology

At present, the CaMV35S constitutive expression promoter is mainly used, and the expression level is low. Moreover, the vaccine antigen with a strong constitutive promoter may affect the growth process of plant cells and inhibit plant growth.

Method used

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  • Plant expression vector for the specific expression of epatitis B surface antigen in fruits

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Embodiment Construction

[0031] A plant expression vector specifically expressing hepatitis B antigen in fruit is characterized in that the promoter in the vector is the promoter of banana lectin gene. The target gene in the vector is the human hepatitis B surface antigen gene. The present invention is described in detail below in conjunction with embodiment:

[0032] 1. Materials

[0033] Banana (MusaAAA group cv. Brazilian) was provided by the Institute of Biotechnology, Chinese Academy of Tropical Agricultural Sciences.

[0034] 2. Method

[0035] (1) Construction of the plant expression vector pCAMBIA2300HBS linked to the HBsAg gene promoter of the lectin gene

[0036] 1. PCR amplification of hepatitis B surface antigen gene

[0037] Design a pair of PCR primers according to the human hepatitis B surface antigen gene HBsAg sequence registered in NCBI:

[0038] HBS1: ACCGAACATGGAGAACAC; HBS2: CGTTTGGTTTTTATTAGGG), using the carrier YEPFLAG-HBS containing the hepatitis B surface antigen gene pr...

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Abstract

The invention relates to a plant expression vector for the specific expression of epatitis B surface antigen (HBsAg) in fruits. The vector adopts the promoter of agglutinin gene expressed specificallyin banana fruits instead of the constitutive 35S promoter on the plant expression vector pBI121, replaces the gus gene on the pBI121 with HBsAg gene and constructs recombinant plant expression vectorpCAMBIA2300HBS. Then agrobacterium infection method is used to transform the vector to transform the banana, PCR detection proves that exogenous gene is inserted in banana genome; and ELISA detectionproves that the promoter of banana lectin gene drives the specific expression of HBsAg gene in fruits. The acquisition of the vector can provide a favorable tool for the specific expression of HBsAggene in fruits and specially lay the foundation of using the vector to prepare hepatitis B edible vaccine. Therefore, the vector has important application value in the future transgenic research.

Description

[technical field] [0001] The invention relates to constructing a plant expression vector by utilizing the banana fruit to specifically express the Banlec gene promoter carrying the hepatitis B virus surface antigen gene. [Background technique] [0002] Banana (Musa spp.) belongs to the Musa family (Musaceae) and is an important tropical and subtropical fruit. Banana meat is sweet and glutinous, fragrant and delicious, and has high nutritional value. In addition to eating fresh and cooking, it can also be processed into a variety of nutritious products. Banana is an ideal plant for producing recombinant medicinal proteins and peptides or valuable compounds by transgenic methods. Its advantages as a bioreactor mainly include the following aspects: 1. Its growth period is short, ten to twelve 2, banana is the fruit with the largest consumption of fresh fruit, fresh food can guarantee that the medicinal protein expressed by the gene in the fruit will not be denatured, and is a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82
Inventor 贾彩红徐碧玉金志强胡伟张建斌刘菊华谭光兰
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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