The design of the constructs of cpsD and simA genes of Streptococcus iniae

oleh: Mohsen Rahmanian Kooshkaki, Mojgan Bandehpour, Bahram Kazemi

Format: Article
Diterbitkan: Shahid Beheshti University of Medical Sciences 2014-04-01

Deskripsi

<p> </p><p class="NBMAbstractText"><span class="NBMAbstractPartsTitlesChar"><span style="line-height: 115%; font-size: 10.5pt;" lang="X-NONE">Background &amp; Objective:</span></span><span lang="X-NONE"> <em>Streptococcosis</em> is one of the bacterial infections in fish, especially rainbow trout which infects brain and nervous systems of fish and is caused by <em>S. iniae</em>. Estimation of the impact of disease prevalence by <em>S. iniae</em> in fish farming in some countries is reported about 100 million dollars per year. Some of the most effective proteins in pathogenicity of these bacteria are <em>SimA</em> and <em>CpsD</em>. In order to design new and effective vaccine, in this study cloning of two genes of Streptococcus was performed into <em>pNZ8148</em> vector and expressed in <em>Escherichia coli</em>. </span></p><p class="NBMAbstractText"><span class="NBMAbstractPartsTitlesChar"><span style="line-height: 115%; font-size: 10.5pt;" lang="X-NONE">Materials and Methods:</span></span><span lang="X-NONE"> <em>simA</em> and <em>cpsD</em> genes were subcloned into <em>pNZ8148</em> vector. Obtained constructs were transformed to expressing <em>E. coli BL21 </em>strain. After induction with nisin, SDS PAGE electrophoresis and Western blotting were used to confirm the procedures.</span></p><p class="NBMAbstractText"><span class="NBMAbstractPartsTitlesChar"><span style="line-height: 115%; font-size: 10.5pt;" lang="X-NONE">Results:</span></span><span lang="X-NONE"> Using PCR with specific and universal primers, the accuracy of cloning was confirmed. Final verification of expressed protein was carried out by SDS-PAGE and western blotting. </span></p><p class="NBMAbstractText"><span class="NBMAbstractPartsTitlesChar"><span style="line-height: 115%; font-size: 10.5pt;" lang="X-NONE">Conclusion:</span></span><span lang="X-NONE"> With regard to the obtained results, it seems that the generated gene construct in this study can be used as a vaccine against <em>Streptococcosis</em> in future researches. </span></p>