TY  - JOUR
T1  - Preparation and Application of Polyclonal Antibody against PKZ from <i>Ctenopharyngodon idellus</i> (CiPKZ)
AU - Hu, Chengyu AU - Fan, Lihua AU - Zhu, Yujiao AU - Tan, Wanlong AU - Yang, Pengjie 
JO  - Journal of Animal and Veterinary Advances
VL  - 11
IS  - 17
SP  - 3169
EP  - 3174
PY  - 2012
DA  - 2001/08/19
SN  - 1680-5593
DO  - javaa.2012.3169.3174
UR  - https://makhillpublications.co/view-article.php?doi=javaa.2012.3169.3174
KW  - PKZ gene
KW  -Za
KW  -polyclonal antibody
KW  -grass carp
KW  -immunohistochemistry
KW  -China
AB  - 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.
ER  -