( Amplification of Epstein-Barr Virus Ex...)Amplification of Epstein-Barr Virus Exon C with Difficult Primers ,,, Using Eppendorf Mastercycler gradient

Amplification of Epstein-Barr Virus Exon C with Difficult Primers
Using Eppendorf Mastercycler gradient

Eng-Lai Tan and Choon-Kook Sam
NPC Laboratory, Institute of Postgraduate Studies and Research, University of Malaya, Kuala Lumpur, Malaysia Eng-Lai Tan Choon-Kook Sam Introduction

Epstein-Barr virus (EBV) is associated with various types of malignancy, in particular, nasopharyngeal carcinoma (NPC). The viral oncogene, latent-membrane protein 1 (LMP-1), is known to transform B-lymphocytes and rodent fibroblast in vitro. It is thought to protect the infected cells from apoptosis by up-regulating the bcl2 and A20 genes.1 LMP-1 gene consists of three exons separated by short introns (Fig. 1).2 One of our investigation efforts attempted to study the possibility of using the LMP-1 protein as an antigenic marker for early progression of NPC.

Fig. 1: Schematic representation of LMP-1 gene. The three exons (EA, EB and EC) are separated by two short introns (I-1 and I-2). Arrows indicate the position of the forward and reverse primers for each exon.

In vitro analysis of LMP-1 requires cloning and expression of this protein for use in antibody detection using ELISA. Each of the three exons of the LMP-1 gene was amplified separately using PCR3 for TA-clonin
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