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Recombinant plasmid with human cytomegalovirus UL140 gene and genetic engineering bacterium and application thereof

A technology of human cytomegalovirus and genetically engineered bacteria, applied in the field of biology, can solve problems such as failure to build successfully

Inactive Publication Date: 2018-07-20
JINAN UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But there is no genetically engineered bacterium containing the UL140 recombinant plasmid that has been successfully constructed

Method used

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  • Recombinant plasmid with human cytomegalovirus UL140 gene and genetic engineering bacterium and application thereof
  • Recombinant plasmid with human cytomegalovirus UL140 gene and genetic engineering bacterium and application thereof
  • Recombinant plasmid with human cytomegalovirus UL140 gene and genetic engineering bacterium and application thereof

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Embodiment Construction

[0033] The present invention will be further described in detail below in conjunction with examples, but the embodiments of the present invention are not limited thereto.

[0034] 1. Plasmid construction

[0035] (1) UL140 gene primer design

[0036] According to the full-length 576bp sequence of the UL140 gene in the TB40 / E (KF297339.1) virus strain genome published on GennBank (the nucleotide sequence is shown in SEQ ID No: 1, and the protein translation product is shown in SEQ ID No: 2), Using this sequence as a template, using the pET32a vector (Youbao Biological Co., Ltd.) as a cloning vector and an expression vector, DNAstar software was used to select appropriate restriction sites for primer design. The designed primers are as follows:

[0037] Table 1 UL140 primer design table

[0038]

[0039] Note: F (upstream primer); R (downstream primer).

[0040] (2) UL140 gene PCR

[0041] According to the gene sequence of HCMV TB40 / E-BAC published on GenBank (GenBank: E...

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Abstract

The invention discloses a recombinant plasmid with human cytomegalovirus UL140 gene and a genetic engineering bacterium and application thereof. The recombinant plasmid with human cytomegalovirus UL140 gene is established by inserting an UL140 gene into an expression type vector pET32a, the recombinant plasmid is further transferred into an expression bacterium BL21, then a genetic engineering bacterium for expressing a poly-histidine tag HCMV UL140 fusion protein can be established, and a corresponding HCMV UL140 fusion protein can be established through expression of the engineering bacterium. The fusion protein disclosed by the invention can be applied to affinity chromatography purification to prepare a polyclonal antibody, then the avoiding mechanism that a chemotactic factor proteinHCMV UL140 avoids a host immune system can be studied, the number of HCMV dormant infection host cells can be reduced, and the immune recognition of a body can be improved.

Description

technical field [0001] The invention relates to the field of biology, in particular to a recombinant plasmid containing human cytomegalovirus UL140 gene, genetically engineered bacteria and applications thereof. Background technique [0002] Cytomegalovirus infection is one of the common viral infections caused by human cytomegalovirus (HCMV). In newborns, immunocompromised and immunosuppressed patients (such as AIDS, organ transplantation or tumors), it can cause serious infection and even death. The pathogenesis of HCMV infection is closely related to the immune function status of the body. During primary HCMV infection, the virus takes mononuclear and polymorphonuclear leukocytes as carriers in human blood and spreads to various organs of the body. [0003] HCMV is extremely widespread in the normal population, rarely causing overt disease, and most people are latently infected. But it poses a great threat to the immunocompromised or damaged population (immunodeficienc...

Claims

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Application Information

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IPC IPC(8): C12N15/70C12N15/38C12N1/21C07K14/045C07K1/22C07K16/08C12R1/19
CPCC07K14/005C07K16/088C12N15/70C12N2710/16122
Inventor 陈风华李瑞林卢杰邢曦雯
Owner JINAN UNIVERSITY
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