@article{MAKHILLJAVA201211173697,
    title = {Preparation and Application of Polyclonal Antibody against PKZ from <i>Ctenopharyngodon idellus</i> (CiPKZ)},
    journal = {Journal of Animal and Veterinary Advances},
    volume = {11},
    number = {17},
    pages = {3169-3174},
    year = {2012},
    issn = {1680-5593},
    doi = {javaa.2012.3169.3174},
    url = {https://makhillpublications.co/view-article.php?issn=1680-5593&doi=javaa.2012.3169.3174},
    author = {Chengyu,Lihua,Yujiao,Wanlong and},
    keywords = {PKZ gene,Za,polyclonal antibody,grass carp,immunohistochemistry,China},
    abstract = {The grass carp (<I>Ctenopharyngodon idellus</I>) PKZ full-length 
  cDNA (GU299765) had been cloned and identified recently. Within its N-terminal 
  part of the protein there are two Z-DNA binding domains called Z&#945;1 (1~67aa) 
  and Z&#945;2 (81~152aa). Z&#945; domain is unique to PKZ and distinguishes them 
  from other translation Initiation Factor 2 alpha (eIF2&#945;)-kinase. To obtain 
  polyclonal antibody against CiPKZ Z&#945;, the <I>PKZ Z&#945;</I> gene was amplified 
  by PCR from the template obtained in the earlier research and identified by 
  DNA sequence analysis. Then, it was digested by BamHI, XhoI and ligated with 
  pET-32a vector which was by the same treatment. Sequenced and blasted with the 
  NCBI GenBank, the recombinant plasmid pET-32a-PKZ Z&#945; was obtained. The 
  recombinant plasmid was transformed into <I>E. coli</I> BL21 (DE3) and induced 
  by 1 mmol L<SUP>-1</SUP> IPTG. Researchers obtained CiPKZ Z&#945; polypeptide 
  via <I>E. coli</I> prokaryotic expression and purified with Ni-NTA His-Bind 
  Resin affinity chromatography. Rabbit Polyclonal Antibody (Pab) against CiPKZ 
  was raised using the purified N-terminal fragment of CiPKZ containing its Z&#945;1 
  and Z&#945;2 domains. Western blot analysis showed that the antibody had high 
  affinity and specificity and higher titer. Immunohistochemistry assay identified 
  that expression of PKZ could be detected in liver tissue.}
    }