@article{MAKHILLJAVA20121133191,
    title = {Cloning of Structural Protein <I>VP1</I> Gene of Foot and Mouth Disease Virus and its Expression in <I>Escherichia coli</I>},
    journal = {Journal of Animal and Veterinary Advances},
    volume = {11},
    number = {3},
    pages = {426-430},
    year = {2012},
    issn = {1680-5593},
    doi = {javaa.2012.426.430},
    url = {https://makhillpublications.co/view-article.php?issn=1680-5593&doi=javaa.2012.426.430},
    author = {Zengqiang,Hongxuan,Hua,Yanying,Guisheng and},
    keywords = {sequence analysis expression,clone,VP1 gene,structrural protein,FMDV,China},
    abstract = {In this study, the viral RNA was extracted from swine FMDV and then the fragment of VP1 was amplified with a primer pair by RT-PCR. The interest fragmen was inserted into pGEM-Teasy vector. There combinant plasmid was identified by restriction analysis and PCR. It was proved by DNA sequencing that the acquired recombinant contains complete <I>VP1</I> gene. The homologie of the nucleotide sequence of <I>VP1 </I>gene were 95.9 and 96.2%, respectively comparing with that of strai O/JPN/00 and O-Tibet-99. Afterwards, the complete <I>VP1</I> gene from the identified recombin an was amplifie with another primer pair containing BamHI and XhoI sites by PCR and digested it with BamHI and XhoI. The expression vector pET28a were digested by BamHI and XhoI, respectively. The target gene <I>VP1</I> was subcloned into vector pET28a. Positive clones named as pET28a-VP1 with interest gene were identificated by restriction analysis, PCR and DNA sequencing. Then there combinant was transformed into <I>Escherichia coli</I> BL21 (DE3) for VP1 expression. The interest gene was induced to express in <I>E. coli </I>with IPTG. The bacteria containing pET28a-VP1 were collected at different time and subsequently were examined by SDS-PAGE and Western-blotting.}
    }