| Abstrak/Abstract |
The development of Animal vaccine is very important to prevent the economical loss in
farming industry. This research aims to develop vector candidate for DNA vaccine based on
chitosan nanoparticles.In this research, plasmid NTC8685 (Nature Technology Corporation),
an antibiotic-free vector, is used as the DNA vaccine vector. Chitosan with medium chain is
used as polymers that entrap the plasmid DNA resulting chitosan-DNA nanoparticles.
Plasmid NTC8685-eGFP was transformed to Escherichia coli NTC4682 as cloning host. The
colony that grow in selection media were confirmed using colony PCR, plasmid extraction,
plasmid PCR and plasmid restriction. The optimization of nanoparticles was done using
different mass ratio of chitosan to plasmid DNA. Ten colonies were chosen randomly and
confirmed using colony-PCR result a single 700 bp band. Plasmid from Clone-10 was
extracted and confirmed using PCR result a single 700 bp band. Plasmid was also confirmed
resulting in two distinct bands: 3000 bp and 700 bp band. The visualization of chitosan-DNA
nanoparticles showed that formulation with mass ratio chitosan: DNA, 1:1,0 can be used to
entrap DNA more efficiently compared to other mass ratios. The mass ratio 1:1,25 or higher
NTC8685-eGFP vector could be applied as vector candidate for DNA vaccine encapsulated
with chitosan nanoparticles. |
