Hai Lian, Kai-Yu Wang, De-Fang Chen, Jun Wang, Ling-Yuan Huang and Cheng-Wei Li
Page: 3800-3805 | Received 21 Sep 2022, Published online: 21 Sep 2022
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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.
Hai Lian, Kai-Yu Wang, De-Fang Chen, Jun Wang, Ling-Yuan Huang and Cheng-Wei Li. Prokaryotic Expression of p1 Gene of Yersinia ruckeri Isolated
from Channel Catfish (Ictalunes punctatus) and Optimization of Expression Conditions.
DOI: https://doi.org/10.36478/javaa.2012.3800.3805
URL: https://www.makhillpublications.co/view-article/1680-5593/javaa.2012.3800.3805