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Recombination human cytomegalovirus fusion protein and its preparing method, application

A technology of human cytomegalovirus and fusion protein, which is applied in the field of preparation of recombinant human cytomegalovirus fusion protein, and can solve the problems of weak antigen specificity and the like

Inactive Publication Date: 2003-07-23
李越希 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Most of the HCMV virus protein antigens used in my country are cultured virus extracts, which contain multiple protein products. The antigen specificity is not strong, and false positives are prone to occur.

Method used

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  • Recombination human cytomegalovirus fusion protein and its preparing method, application
  • Recombination human cytomegalovirus fusion protein and its preparing method, application
  • Recombination human cytomegalovirus fusion protein and its preparing method, application

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Embodiment Construction

[0068] Detailed description of the embodiment of the present invention:

[0069] 25 Amino Acids of C-terminus of Human Cytomegalovirus PP150 Protein and gp52 Protein

[0070] Fusion expression of C-terminal 164 amino acids

[0071] A gene fragment of 25 amino acids at the C-terminus of the PP150 protein was chemically synthesized by using codons favored by bacteria. The gp52 protein gene fragment was amplified by PCR method. The two gene fragments were cloned into the NcoI / BamHI and BamHI / EcoRI sites in the same plasmid pET28a(+), so that the two were connected in series, and the translation frame was consistent, and a fusion protein could be expressed. The recombinant plasmid was transformed into Escherichia coli BL21 (DE3), and the engineered bacteria that highly expressed the fusion protein was obtained through screening. The expressed fusion protein accounted for about 35% of the total protein in the bacteria and existed in a soluble form....

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Abstract

A novel human cytomegalovirus fusion protein used for detecting human cytomegalovirus antibody or antigen and preparing monoclonal or polyclonal antibody is prepared by recombining human cytomegalvirus PP150 protein with gp52 protein in such manner that the 25 amino acids at c terminal of pp150 and the 164 amino acids at C terminal of gp52 are serially linked via 2 amino acids Gly and Ser.

Description

technical field [0001] The invention utilizes genetic engineering technology to prepare recombinant human cytomegalovirus fusion protein. The two fragments of the fusion protein come from the gp52 protein and PP150 protein of human cytomegalovirus respectively. The two fragments are linked and expressed by genetic engineering technology to form a new fusion protein. The new fusion protein produced can be used as an antigen for human cytomegalovirus. Detection of cell virus antibody or antigen, etc. The invention relates to the fields of genetic engineering technology and diagnostic reagents. technical background [0002] Human cytomegalovirus (HCMV for short) is a DNA virus that can remain latent in the body for a long time after infecting the human body, and can be activated to produce recurrent infection. HCMV-infected pregnant women can not only infect the fetus through the placenta, but also infect the newborn through the birth canal and breast milk. After the fetus an...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/09C12N15/12G01N33/68
Inventor 李越希陶开华
Owner 李越希
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