Cloning of vacll into pNMN013: development of a recombinant BCG vaccine against tuberculosis
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Date
2004
Authors
Ramli, Siti Nur Rasyidah Md
Journal Title
Journal ISSN
Volume Title
Publisher
Pusat Pengajian Sains Kesihatan, Universiti Sains Malaysia
Abstract
In previous study, the Vacll gene has been constructed through the
technique of assembly PCR and has been cloned into pKK to produce surface
display vaccine candidate (pTMSinakVacll). Vacll gene is a synthetic gene
consists of selected T cell epitopes of Mycobacterium tuberculosis (MTB) genes
namely ESAT6, MTP40, 38kD and MPT64.
In this study, Vacll gene was cloned into pNMN013 for development of a
recombinant BCG vaccine. PCR was used for the amplification of the gene DNA
from pTMSinakVacll. The PCR products (Vacll gene) were visualized directly on
agarose gel. The PCR product was first cloned into pTOPO vector. The PCR
product were also digested with Nhel and Ndel and cloned into Nhel and Ndel
digested pNMN013. The plasmids were transformed into E.coli. There were
some colonies grown on the agar plate.
Finally, the colonies were screened using PCR technique and restriction
enzyme analysis. But after the screening procedures were done no expected
recombinant plasmid were obtained.
Description
Keywords
vaccine