Prokaryotic Expression of p1 Gene of Yersinia ruckeri Isolated from Channel Catfish (Ictalunes punctatus) and Optimization of Expression Conditions

被引：0

作者：

Lian, Hai ^{[1
,3
]}

Wang, Kai-Yu ^{[1
,3
]}

Chen, De-Fang ^{[2
,3
]}

Wang, Jun ^{[1
,3
]}

Huang, Ling-Yuan ^{[1
,3
]}

Li, Cheng-Wei ^{[1
,3
]}

机构：

[1] Sichuan Agr Univ, Coll Vet Med, Dept Basic Vet, Yaan 625014, Sichuan, Peoples R China

[2] Sichuan Agr Univ, Coll Anim Sci & Technol, Dept Aquaculture, Yaan 625014, Sichuan, Peoples R China

[3] Sichuan Agr Univ, Key Lab Anim Dis & Human Hlth Sichuan Prov, Yaan 625014, Sichuan, Peoples R China

来源：

JOURNAL OF ANIMAL AND VETERINARY ADVANCES | 2012年 / 11卷 / 20期

关键词：

Yersinia ruckeri; p1; gene; prokaryotic expression; optimization; China; RNA-POLYMERASE; CLONED GENES; BACTERIUM; PROTEASE; DISEASE;

D O I：

暂无

中图分类号：

S85 [动物医学（兽医学）];

学科分类号：

0906 ;

摘要：

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 degrees C for 4 h with 0.8 mM IPTG.

引用

页码：3800 / 3805

页数：6

共 15 条

[1] Bo-Hai X, 1991, CHINESE SCI BULL, V8, P620
[2] THE SIGNAL FOR THE TERMINATION OF PROTEIN-SYNTHESIS IN PROCARYOTES
BROWN, CM
STOCKWELL, PA
TROTMAN, CNA
TATE, WP
[J]. NUCLEIC ACIDS RESEARCH, 1990, 18 (08) : 2079 - 2086
[3] Fang-Ling F, 2010, OCEANOLOGIA LIMNOLOG, V41, P862
[4] In vitro and in vivo studies of the Yrp1 protease from Yersinia ruckeri and its role in protective immunity against enteric red mouth disease of salmonids
Fernandez, L
Lopez, JR
Secades, P
Menendez, A
Marquez, I
Guijarro, JA
[J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2003, 69 (12) : 7328 - 7335
[5] The iron- and temperature-regulated haemolysin YhIA is a virulence factor of Yersinia ruckeri
Fernandez, Lucia
Prieto, Miguel
Guijarro, Jose A.
[J]. MICROBIOLOGY-SGM, 2007, 153 : 483 - 489
[6] Furones M. D., 1993, Annual Review of Fish Diseases, V3, P105, DOI 10.1016/0959-8030(93)90031-6
[7] Mastering seeds for genomic size nucleotide BLAST searches
Gotea, V
Veeramachaneni, V
Makalowski, W
[J]. NUCLEIC ACIDS RESEARCH, 2003, 31 (23) : 6935 - 6941
[8] MIERENDORF R, 1994, INNOVATIONS, V1, P1
[9] ROMALDE JL, 1993, FEMS MICROBIOL LETT, V112, P291, DOI 10.1111/j.1574-6968.1993.tb06465.x
[10] DESCRIPTION OF A BACTERIUM ASSOCIATED WITH REDMOUTH DISEASE OF RAINBOW TROUT(SALMO GAIRDNERI)
ROSS, AJ
RUCKER, RR
EWING, WH
[J]. CANADIAN JOURNAL OF MICROBIOLOGY, 1966, 12 (04) : 763 - &

← 1 2 →