A description of the pFastBACquad constructs prepared in this study from genomic data obtained from human stool samples containing human African rotavirus strains.

<p>All codon-optimised ORFs were inserted into a commercial pUC57 transport plasmid at GeneArt (Life Technologies, New York, NY) and GenScript (GenScript USA Inc. New Jersey, NJ).</p>a<p><b>pFBqG2, pFBqG8 and pFBqG12</b>: Generated by ligating coding regions for G2, G8 and G12 VP7 proteins excised from pUC57G2, pUC57G2 and pUC57G8 plasmids to pFBq vector DNA, respectively. Both insert and vector DNA were prepared by double-digestion with <i>Bam</i> HI and <i>Not</i> I.</p>¥<p>VP4 and VP7 outer capsid proteins expressed by baculoviruses prepared from these expression cassettes were used to generate RV-VLPs in the current study. The VP2 and VP6 proteins that were prepared from strain RVA/Human-wt/ZAF/GR10924/1999/G9P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Madhi1" target="_blank">[6]</a> formed the scaffolds on to which the outer capsid proteins were assembled.</p>b<p><b>pFBqP</b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a><b> and pFBqP</b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a>: Generated by ligating coding regions for P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a> and P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a> VP4 proteins excised from pUC57P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a> and pUC57P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a> plasmids to pFBq vector DNA. The vector and insert DNA were prepared by digestion with <i>Sma</i> I and <i>Spe</i> I.</p>c<p><b>pFBqG2P</b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a><b>, pFBqG2P</b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a>: Generated by ligating the coding regions for P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a> and P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a> VP4 proteins excised from pUC57P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a> and pUC57P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a> plasmids to recombinant pFBqG2 expression cassettes, respectively. Both insert and vector DNA were prepared by double-digestion with <i>Sma</i> I and <i>Spe</i> I.</p>d<p><b>pFBqG8P</b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a><b>, pFBqG8P</b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a>: Generated by ligating coding regions for P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a> and P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a> VP4 proteins excised with <i>Sma</i> I and <i>Spe</i> I from pUC57P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a> and pUC57P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a> plasmids to the recombinant pFBqG8 expression cassettes, respectively. The vector was also prepared by digesting with <i>Sma</i> I and <i>Spe</i> I.</p>e<p><b>pFBqG12P</b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a><b>, pFBqG12P</b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a>: Generated by ligating the coding regions for P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a> and P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a> VP4 proteins excised from pUC57P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a> and pUC57P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a> plasmids to the recombinant pFBqG12 expression vector. Both the insert and vector were double-digested with <i>Sma</i> I and <i>Spe</i> I.</p>f<p><b>FBqG9P</b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a><b>, pFBqG9P</b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a><b>, pFBqG2P</b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Madhi1" target="_blank">[6]</a><b>, pFBqG8P</b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Madhi1" target="_blank">[6]</a><b>, pFBqG12P</b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Madhi1" target="_blank">[6]</a>: Double-digestion of pFBqG9P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Madhi1" target="_blank">[6]</a> with <i>Sma</i> I and <i>Spe</i> I resulted in pFBqG9_<i>Sma</i>I/<i>Spe</i>I and P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Madhi1" target="_blank">[6]</a>_<i>Sma</i>I/<i>Spe</i>I DNA fragments. Double digesting pFBqG9P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Madhi1" target="_blank">[6]</a> with <i>Bam</i> HI and <i>Not</i> I resulted in pFBqP<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Madhi1" target="_blank">[6]</a><i>_Bam</i>HI/<i>Not</i>I and G9_<i>Bam</i>HI/<i>Not</i>I fragments. Recombinant pFBqG9P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a>, pFBqG9P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a>, pFBqG2P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Madhi1" target="_blank">[6]</a> expression cassettes were engineered by ligating coding regions for P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Vesikari1" target="_blank">[4]</a> and P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Glass1" target="_blank">[8]</a> VP4 proteins to the recombinant pFBqG9_<i>Sma</i>I/<i>Spe</i>I vector. Ligating G2_<i>Bam</i>HI/<i>Not</i>I, G8_<i>Bam</i>HI/<i>Not</i>I and G12_<i>Bam</i>HI/<i>Not</i>I fragments to recombinant pFBqP<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Madhi1" target="_blank">[6]</a>_<i>Bam</i>HI/<i>Not</i>I vector resulted in pFBqG2P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Madhi1" target="_blank">[6]</a>, pFBqG8P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Madhi1" target="_blank">[6]</a>, pFBqG12P<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105167#pone.0105167-Madhi1" target="_blank">[6]</a> expression cassettes, respectively.</p><p>A description of the pFastBACquad constructs prepared in this study from genomic data obtained from human stool samples containing human African rotavirus strains.</p>