Construction and in vitro characterization of a molecularly cloned human immunodeficiency virus type 1 library

Tetsu Mukai, Takeshi Kurosu, Masanobu Kinomoto, Satoshi Komoto, Miki Shiraga, Wattana Auwanit, Kazuyoshi Ikuta

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Development of a safe and effective vaccine against human immunodeficiency virus (HIV) is urgent, but many concerns regarding the safety and efficacy of the currently developing vaccines remain. A major hindrance in HIV vaccine development is the genetic diversity, a hallmark of HIV biology, and a poor understanding of how HIV vaccine prevents the emergence of escape variants during infection and progression of AIDS. Here, we developed a method to construct a molecularly cloned viral library. This technique employs a long-range polymerase chain reaction (PCR) to amplify a virtually full-length HIV type 1 (HIV-1) provirus genome from peripheral blood mononuclear cells (PBMCs) infected with CRF01_AE (subtype E) Thai primary isolate. Among randomly selected 93 clones, 41 with a full-length sequence were able to replicate in PBMCs, 5 of which induced strong cytopathic effects. Replication kinetics also showed that the parental virus was intermediate among the clones. Thus, the molecular library prepared by this method showed the quasi-species in infected cells and this method could provide a new possibility for the development of an order-made therapeutic vaccine against HIV-1.

Original languageEnglish
Pages (from-to)1181-1185
Number of pages5
JournalVaccine
Volume20
Issue number7-8
DOIs
Publication statusPublished - 15-01-2002

Fingerprint

Human immunodeficiency virus
Human immunodeficiency virus 1
HIV-1
Vaccines
vaccines
HIV
mononuclear leukocytes
clones
proviruses
Blood Cells
Clone Cells
cytopathogenicity
vaccine development
Proviruses
methodology
polymerase chain reaction
Libraries
kinetics
Acquired Immunodeficiency Syndrome
Biological Sciences

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Immunology and Microbiology(all)
  • veterinary(all)
  • Public Health, Environmental and Occupational Health
  • Infectious Diseases

Cite this

Mukai, Tetsu ; Kurosu, Takeshi ; Kinomoto, Masanobu ; Komoto, Satoshi ; Shiraga, Miki ; Auwanit, Wattana ; Ikuta, Kazuyoshi. / Construction and in vitro characterization of a molecularly cloned human immunodeficiency virus type 1 library. In: Vaccine. 2002 ; Vol. 20, No. 7-8. pp. 1181-1185.
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Construction and in vitro characterization of a molecularly cloned human immunodeficiency virus type 1 library. / Mukai, Tetsu; Kurosu, Takeshi; Kinomoto, Masanobu; Komoto, Satoshi; Shiraga, Miki; Auwanit, Wattana; Ikuta, Kazuyoshi.

In: Vaccine, Vol. 20, No. 7-8, 15.01.2002, p. 1181-1185.

Research output: Contribution to journalArticle

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