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

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.

journal_name

Vaccine

journal_title

Vaccine

authors

Mukai T,Kurosu T,Kinomoto M,Komoto S,Shiraga M,Auwanit W,Ikuta K

doi

10.1016/s0264-410x(01)00429-7

subject

Has Abstract

pub_date

2002-01-15 00:00:00

pages

1181-5

issue

7-8

eissn

0264-410X

issn

1873-2518

pii

S0264410X01004297

journal_volume

20

pub_type

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