NDLI: Functionally active rat S100A4 from a polymerase chain reaction-synthesized gene expressed in soluble form in Escherichia coli

Content Provider	PubMed Central
Author	Liu, Ziquan Xu, Chuanxiang Zhang, Jianwei Chen, Yunyun Liu, Xiaohua Lei, Wu Zhang, Zhiqing Meng, Xiangyan Liu, Hongtao Jiang, Zifeng Wang, Tianhui
Copyright Year	2014
Abstract	S100A4 protein is associated with Ca2+-dependent regulation of intracellular activities and is significant in the invasion, growth and metastasis of cancer. In order to express rat S100A4 functionally and identify its biological activity following purification, an S100A4 gene fragment was optimized and fully synthesized via overlapping polymerase chain reaction. The gene was inserted into the prokaryotic expression vector, pBV220, with phage λ PRPL promoters following confirmation by DNA sequencing. The pBV220-S100A4 plasmid was constructed and transformed into Escherichia coli DH5α. Following temperature induction, rat S100A4 was overexpressed and the protein was observed to be located in the supernatant of the lysates, which was ~30–40% of the total protein within the host. The protein was isolated and purified by metal-chelate affinity chromatography. High purity protein (>98% purity) was obtained and in vitro western blot analysis identified that the recombinant S100A4 was able to bind to the antibody against wild-type S100A4. The bioactivity of the recombinant protein was detected via Transwell migration and invasion assays. The polyclonal antibody of rat S100A4 protein was prepared for rabbit immunization and exhibited similar efficacies when compared with commercial S100A4. Therefore, rat S100A4 was functionally expressed in E. coli; thus, the production of active recombinant S100A4 protein in E. coli may further aid with the investigation and application of S100A4.
Related Links	http://dx.doi.org/10.3892/ol.2014.1870
Ending Page	1184
Page Count	6
Starting Page	1179
File Format	PDF
ISSN	17921074
e-ISSN	17921082
Journal	Oncology Letters
Issue Number	4
Volume Number	7
Language	English
Publisher	D.A. Spandidos
Publisher Date	2014-02-01
Access Restriction	Open
Rights Holder	D.A. Spandidos
Subject Keyword	Cancer Research Oncology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Cancer Research Oncology

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Detection of the eaeA Gene in Escherichia coli from Chickens by Polymerase Chain Reaction

Detection of the eaeA Gene in Escherichia coli from Chickens by Polymerase Chain Reaction

Detection of the eaeA Gene in Escherichia coli from Chickens by Polymerase Chain Reaction

Streptococcus pneumoniae polA gene is expressed in Escherichia coli and can functionally substitute for the E. coli polA gene.

A Novel Form of RNA Polymerase from Escherichia coli

Detection of enterotoxigenic Escherichia coli after polymerase chain reaction amplification with a thermostable DNA polymerase.

Characterization of Soluble Hepatitis C Virus RNA-Dependent RNA Polymerase Expressed in Escherichia coli

Total Reconstitution of Functionally Active 50S Ribosomal Subunits from Escherichia coli *

GENOMIC FINGERPRINTING OF ESCHERICHIA COLI STRAINS USING REPETITIVE SEQUENCE BASED POLYMERASE CHAIN REACTION

Functionally active rat S100A4 from a polymerase chain reaction-synthesized gene expressed in soluble form in Escherichia coli

Similar Documents

Detection of the eaeA Gene in Escherichia coli from Chickens by Polymerase Chain Reaction

Detection of the eaeA Gene in Escherichia coli from Chickens by Polymerase Chain Reaction

Detection of the eaeA Gene in Escherichia coli from Chickens by Polymerase Chain Reaction

Streptococcus pneumoniae polA gene is expressed in Escherichia coli and can functionally substitute for the E. coli polA gene.

A Novel Form of RNA Polymerase from Escherichia coli

Detection of enterotoxigenic Escherichia coli after polymerase chain reaction amplification with a thermostable DNA polymerase.

Characterization of Soluble Hepatitis C Virus RNA-Dependent RNA Polymerase Expressed in Escherichia coli

Total Reconstitution of Functionally Active 50S Ribosomal Subunits from Escherichia coli *

GENOMIC FINGERPRINTING OF ESCHERICHIA COLI STRAINS USING REPETITIVE SEQUENCE BASED POLYMERASE CHAIN REACTION

Functionally active rat S100A4 from a polymerase chain reaction-synthesized gene expressed in soluble form in Escherichia coli