PMID- 29596634
OWN - NLM
STAT- MEDLINE
DCOM- 20190905
LR  - 20190906
IS  - 1574-6968 (Electronic)
IS  - 0378-1097 (Linking)
VI  - 365
IP  - 9
DP  - 2018 May 1
TI  - Expression and purification of biologically active recombinant rabbit monocyte 
      chemoattractant protein1 in Escherichia coli.
LID - 10.1093/femsle/fny070 [doi]
AB  - Monocyte chemoattractant protein 1 (MCP1) with recruiting monocytes is an 
      important factor at the beginning of inflammatory disorders such as 
      atherosclerosis which seems its blocking preclude this process and help 
      improvement of related diseases. To perform clinical research in this field, MCP1 
      protein is required but firstly, animal studies should be done. As the rabbit is 
      a suitable model for many inflammatory disorders, and Escherichia coli BL21(DE3) 
      (BL21) cell is a high-efficiency host for protein expression, we decided to 
      produce recombinant rabbit MCP1 (rRMCP1) in BL21/pET28a system. After codon 
      usage, a construct containing RMCP1 sequence was synthesized, cloned into the 
      pET28a plasmid, and overexpressed in BL21 cells. Followed that, with changing 
      expression condition such as cell concentration before the induction, time 
      period, temperature, shaking rate and inducer concentration (IPTG), rRMCP1 
      expression was optimized, and purified by Ni-NTA. The biological activity of the 
      expressed protein was verified using monocyte migration assay. Using this 
      expression system, nearly 28 mg/mL rRMCP1 was produced at 26 degrees C/180 rpm for 24 h 
      in LB broth medium with 1 mM IPTG. Therefore, we were succeeded to express the 
      intermediate level of rRMCP1 with this method. This amount of protein is 
      sufficient for biological researches in the laboratory.
FAU - Boshtam, Maryam
AU  - Boshtam M
AD  - Isfahan Cardiovascular Research Center, Cardiovascular Research Institute, 
      Isfahan University of Medical Sciences, Isfahan 8174643446, Iran.
FAU - Khanahmad Shahreza, Hossein
AU  - Khanahmad Shahreza H
AD  - Genetic and Molecular Biology, Faculty of Medicine, Isfahan University of Medical 
      Sciences, Isfahan 8174643446, Iran.
FAU - Feizollahzadeh, Sadegh
AU  - Feizollahzadeh S
AD  - Faculty of Paramedical, Urmia University of Medical Sciences, Urmia 5756115198, 
      Iran.
FAU - Rahimmanesh, Ilnaz
AU  - Rahimmanesh I
AD  - Genetic and Molecular Biology, Faculty of Medicine, Isfahan University of Medical 
      Sciences, Isfahan 8174643446, Iran.
FAU - Asgary, Sedigheh
AU  - Asgary S
AD  - Isfahan Cardiovascular Research Center, Cardiovascular Research Institute, 
      Isfahan University of Medical Sciences, Isfahan 8174643446, Iran.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - FEMS Microbiol Lett
JT  - FEMS microbiology letters
JID - 7705721
RN  - 0 (Chemokine CCL2)
RN  - 0 (Codon)
RN  - 0 (Recombinant Proteins)
SB  - IM
MH  - Animals
MH  - Chemokine CCL2/*genetics/*isolation & purification/metabolism
MH  - Cloning, Molecular
MH  - Codon/genetics
MH  - Escherichia coli/*genetics/metabolism
MH  - *Gene Expression
MH  - Protein Engineering
MH  - Rabbits/genetics
MH  - Recombinant Proteins/genetics/isolation & purification/metabolism
EDAT- 2018/03/30 06:00
MHDA- 2019/09/07 06:00
CRDT- 2018/03/30 06:00
PHST- 2018/01/29 00:00 [received]
PHST- 2018/03/26 00:00 [accepted]
PHST- 2018/03/30 06:00 [pubmed]
PHST- 2019/09/07 06:00 [medline]
PHST- 2018/03/30 06:00 [entrez]
AID - 4955552 [pii]
AID - 10.1093/femsle/fny070 [doi]
PST - ppublish
SO  - FEMS Microbiol Lett. 2018 May 1;365(9). doi: 10.1093/femsle/fny070.