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Modifiable loca of human type 3 adenovirus hexon and application thereof

An adenovirus and hexon technology, applied in the field of molecular biology, can solve problems such as application limitations, large differences in the sequence and length of the HVR region, and research on whether amino acids can be replaced

Inactive Publication Date: 2011-05-18
GUANGZHOU INST OF RESPIRATORY DISEASE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

), but the main purpose of this study is whether the HVR region of adenovirus can be modified, rather than how to modify, so this study did not study whether certain amino acids in HVR can be replaced or not, and human adenovirus type 5 is in In genotyping, it belongs to group C (HAdV-C), while human adenovirus type 3 belongs to group B (HAdV-B). The sequences and lengths of the HVR regions of the two are quite different
Moreover, due to the widespread presence of neutralizing antibodies against human adenovirus type 5 in the population, the application of expressing recombinant vaccines and therapeutic drugs using it as a vector is greatly restricted.

Method used

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  • Modifiable loca of human type 3 adenovirus hexon and application thereof
  • Modifiable loca of human type 3 adenovirus hexon and application thereof
  • Modifiable loca of human type 3 adenovirus hexon and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: The first mutation experiment of PBRADV3ΔE3EGFP hexon hypervariable region

[0033]The hexon hypervariable regions HVR1, HVR2, HVR5, and HVR7 of wild-type HADV7 were replaced with the corresponding positions of the hexon hypervariable regions on the viral vector PBRADV3ΔE3EGFP (shown underlined in the amino acid sequence), and the mutated amino acid sequences are shown in the underlined part in Table 1 shown.

[0034] Table 1

[0035]

HADV7

HADV3

HVR1

IVTT GE DNATTY T

IVTT NRDNAVTTTTTN T

HVR2

KEGL EIGKDITADN KPIYADK

KEGL QIGKDITTTEGEE KPIYADK

HVR5

DGR EAADAFS PEIVLYTEN

DGR DAVAGALA PEIVLYTEN

[0036] HVR7

IK PRDTA WEKDT

IK VKTDDANG WEKDAN

[0037] The primer sequences used are shown in Table 2, where the underlines in the HexU and HexD sequences indicate the restriction sites of Cla I and BamH I, respectively.

[0038] Table 2

[0039]

...

Embodiment 2

[0042] Example 2: The second mutation experiment of PBRADV3ΔE3EGFP hexon hypervariable region

[0043] The hexon hypervariable regions HVR1, HVR2, HVR5, and HVR7 of wild-type HADV4 were respectively replaced with the corresponding positions of the hexon hypervariable regions on the viral vector PBRADV3ΔE3EGFP (shown underlined in the amino acid sequence), and the mutated amino acid sequences are shown in the underlined part in Table 3 shown.

[0044] table 3

[0045]

HADV4

HADV3

HVR1

KDSDSKM

IVTTNRDNAVTTTTNT

HVR2

EADGLPIRIDSTSGTDTVIYADK

KEGLOIGKDITTTEGEEKPIYADK

HVR5

D. SKTIVANYDPDIVM YTEN

DGRDAVAGALAPEIVLYTEN

HVR7

VKVKTDAGSEKWDKDDTTV

IKVKTDDANGWEKDAN

[0046] The primer sequences used are shown in Table 4, wherein the underlines in the HexU and HexD sequences indicate the restriction sites of Cla I and BamH I, respectively.

[0047] Table 4

[0048]

[0049] The...

Embodiment 3

[0050] Example 3: The third mutation experiment of PBRADV3ΔE3EGFP hexon hypervariable region

[0051] The hexon hypervariable regions HVR1, HVR2, HVR5, and HVR7 of wild-type HADV4 were respectively replaced with the corresponding positions of the hexon hypervariable regions on the viral vector PBRADV3ΔE3EGFP (shown underlined in the amino acid sequence), and the mutated amino acid sequence is shown in the underlined part in Table 5 shown.

[0052] table 5

[0053]

HADV4

HADV3

HVR1

KDSDSKM

IVTT NRDNAVTTTTTN T

HVR2

EADGL PIRIDSTSGTDTV IYADK

KEGLQIGKDIT TTEGEE KPIYADK

HVR5

D. SKTIVANYDPD IVMYTEN

DGR DAVAGALA PEIVLYTEN

HVR7

VKVKTD AGSEK WDKDDTTV

IKVKTD DANG WEKDAN

[0054] The primer sequences used are shown in Table 6, wherein the underlines in the HexU and HexD sequences indicate the restriction sites of Cla I and BamH I, respectively.

[0055] Table 6

[0056]

[0...

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Abstract

The invention provides modifiable amino acid loca in hypervariable regions of human type 3 adenovirus hexon. The modifiable loca are positioned in the hypervariable regions of the hexon and have at least one amino acid locus in a sequence shown as SEQ ID NO:1-4. The amino acid loca are positioned in hypervariable regions 1, 2, 5, and 7 of the human type 3 adenovirus hexon. The invention also provides the application of the modifiable amino acid loca to the show of extrinsic protein and the preparation of immunogen, medicaments and detection reagents. The modifiable amino acid loca lay the foundation for a virus show technology platform which is researched and developed by using a technology platform of modifiable extrinsic polypeptides in the hypervariable regions of the human type 3 adenovirus hexon as vaccines or antigens.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to the modifiable site of human type 3 adenovirus and its application. Background technique [0002] Human adenovirus (Adenovirus) was isolated and identified from patients with acute respiratory diseases in 1953. It is a kind of pathogen that mainly infects the human respiratory tract and digestive tract. So far, humans have discovered 6 species of adenoviruses with 52 different serotypes. Adenoviruses have a wide range of prevalence and fast transmission speed. There is a great threat. Epidemiological surveys show that in my country, the subtypes of adenoviruses that cause severe respiratory infections are mainly types 3, 4, and 7, and type 11 is relatively rare. [0003] The DNA of adenovirus is double-stranded linear, with a total length of 30-38kb. Foreign genes can be inserted into the E1 and E3 regions encoding early proteins for expression, and research on recombinant vac...

Claims

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Application Information

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IPC IPC(8): C07K14/075C07K7/08A61K39/235A61K38/16A61K38/10G01N33/68G01N33/569C12N7/01C12N15/861
Inventor 周荣李潇李海涛高文娟田新贵钟南山
Owner GUANGZHOU INST OF RESPIRATORY DISEASE
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