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Malinee Saelim. Generation and characterization of chimeric DENV-1/2 virus-like particles. Master's Degree(Microbiology). Chiang Mai University Library. : Chiang Mai University, 2016.
Generation and characterization of chimeric DENV-1/2 virus-like particles
การสร้างและศึกษาคุณสมบัติของอนุภาคคล้ายไวรัสลูกผสมระหว่างเชื้อไวรัสเด็งกี่ ชนิดที่ 1 และ ชนิดที่ 2
Name: Malinee Saelim
ThaSH:
Dengue viruses
Classification :.DDC: 572.6
Abstract:
Dengue virus (DENV) infection causes undifferentiated fever, dengue fever, dengue
hemorrhagic fever, and dengue shock syndrome. The first license for use of dengue
vaccine in human involves a live-attenuated vaccine based on yellow fever virus 17D in
which the prM and E coding region was substituted with those of the four serotypes of
dengue virus. This vaccine could not protect diseases caused by DENV-1 and -2
infections with high efficacy in children younger than 9 year olds. Virus-like particles
have been proposed as an alternative vaccine candidate as they resemble virus particles
in structure but lack the viral genome, which is required for virus replication and
pathogenesis, resulting in greater safety profile. In this study, chimeric DENV-1/2 viruslike particles were generated by transfecting C6/36 cells with an insect cell expression
plasmid encoding the prM and E genes of DENV-2 in which the domain III and the stemanchor region of E protein had been substituted with corresponding regions of DENV-1
and Japanese encephalitis virus (JEV), respectively. Selection with blasticidin and
subsequent cloning attempts resulted in transfected cell clones with high level of
expression of E protein in the cytoplasm. SDS-PAGE analysis of partially purified
extracellular E-containing materials revealed three structural protein bands (E, prM and
M) and an unknown protein band. The presence of prM and E was confirmed by
employing specific monoclonal antibodies in a western blot analysis. Transmission
electron microscopic analysis revealed spherical particles with heterogeneity in size,
compatible with the presence of viral particles that lack internal core structure.
Cleavage of prM in these particles was reduced from that of the parental DENV-2
virus-like particles. These results indicate that the DENV-2 prM protein and chimericDENV-2/1/JEV E protein can assemble into spherical particles of various class sizes for
release from transfected C6/36 cells.
Chiang Mai University Library
Address:
CHIANG MAI
Email:
cmulibref@cmu.ac.th
Name: Arunee Thitithanyanont
Role:
Chairperson
Name: Nopporn Sittisombut
Role:
Member
Name: Poonsook Keelapang
Role:
Member
Created:
2016
Modified:
2018-11-04
Issued:
2018-11-04
วิทยานิพนธ์/Thesis
application/pdf
CallNumber:
Th 572.6 M251G
eng
DegreeName: Master of Science
Level: Master's Degree
Descipline: Microbiology
Grantor: Chiang Mai University
ใช้เวลา
0.01881 วินาที
Malinee Saelim
| Title | Contributor | Type |
|---|---|---|
| Generation and characterization of chimeric DENV-1/2 virus-like particles
มหาวิทยาลัยเชียงใหม่ Malinee Saelim | Arunee Thitithanyanont Nopporn Sittisombut Poonsook Keelapang | วิทยานิพนธ์/Thesis |
Arunee Thitithanyanont
Nopporn Sittisombut
Poonsook Keelapang