TY - JOUR
T1 - Prokaryotic Expression of p1 Gene of Yersinia ruckeri Isolated
from Channel Catfish (Ictalunes punctatus) and Optimization of Expression Conditions
AU - Wang, Kai-Yu AU - Lian, Hai AU - Chen, De-Fang AU - Wang, Jun AU - Huang, Ling-Yuan AU - Li, Cheng-Wei
JO - Journal of Animal and Veterinary Advances
VL - 11
IS - 20
SP - 3800
EP - 3805
PY - 2012
DA - 2001/08/19
SN - 1680-5593
DO - javaa.2012.3800.3805
UR - https://makhillpublications.co/view-article.php?doi=javaa.2012.3800.3805
KW - Yersinia ruckeri
KW -p1 gene
KW -prokaryotic expression
KW -optimization
KW -China
AB - The p1 gene of Yersinia ruckeri (yrp1)
which was isolated from channel catfish was amplified by PCR with specific primers
and inserted into pMD19-T vector. The positive recombinant plasmid was selected
and sequenced. Then, the yrp1 gene was subcloned into pET-32a (+) vector
and transformed into BL21 (DE3) followed by induction with IPTG and detection
with SDS-PAGE. Optimization of the induction conditions were conducted. The
results showed that the recombinant protein with a molecular mass of about 72
kDa was mostly packaged into inclusion bodies. The optimization of induction
process conditions led us to perform the fusion protein induction at 37°C
for 4 h with 0.8 mM IPTG.
ER -