NDLI: An improved strategy for efficient expression and purification of soluble HIV-1 tat protein in E.coli

Content Provider	Springer Nature Link
Author	Zhang, Shi meng Fan, Rong Yang, Tian yi Sun, Yi Li, Jing yun Xu, Qin zhi Zhou, Ping kun
Copyright Year	2009
Abstract	Although the endogenous function of Tat has been elucidated in the past twenty years, the study of its exogenous activity has been hampered due to the difficulty of large scale preparation of the active Tat protein. To express the full-length Tat protein in E.coli, the tat gene was cloned from an HIV infected patient by overlapping PCR. Rare codon usage analysis showed that rare E.coli codons, especially consecutive rare codons for Arg, account for 14% (14 of 101) rare E.coli codons in the tat gene. The expression of the HIV-1 tat gene was verified to be very poor in strain BL21 (DE3) due to the abundance of rare codons; however, tat gene expression was found to be very efficient in the host strain of Rosetta-gami B (DE3), which was supplemented with six rare tRNAs for Arg, Leu, Ile and Pro. Subsequent purification revealed that the proteins are soluble and unusually, the tagged Tat can form dimers independent of cystine disulfide bonds. The purity, integrity and molecular weight of the Tat protein were demonstrated by MALDI-TOF mass spectrometry. Reporter gene activating assay was further confirmed by investigating the transactivation activity of the recombinant Tat protein. Our improved strategy for efficient high level expression and purification of soluble Tat protein has paved the way to fully investigate its exogenous function.
Starting Page	518
Ending Page	528
Page Count	11
File Format	PDF
ISSN	16740769
Journal	Virologica Sinica
Volume Number	24
Issue Number	6
e-ISSN	1995820X
Language	English
Publisher	SP Wuhan Institute of Virology, CAS
Publisher Date	2009-11-27
Publisher Place	Heidelberg
Access Restriction	Subscribed
Subject Keyword	HIV tat gene E.coli Protein expression Codon usage Microbial Genetics and Genomics Microbiology Biochemistry Oncology Medical Microbiology Virology
Content Type	Text
Resource Type	Article
Subject	Infectious Diseases Virology Immunology Molecular Medicine

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

Characterization of synonymous codon usage bias in the pseudorabies virus US1 gene

Analysis of synonymous codon usage bias in 09H1N1

A new strategy for full-length Ebola virus glycoprotein expression in E.coli

Prokaryotic expression and purification of HIV-1 Vif and hAPOBEC3G, preparation of polyclonal antibodies

Synonymous codon usage bias and overexpression of a synthetic gene encoding Interferon α2b in yeast

Efficient gene transfer mediated by HIV-1-based defective lentivector and inhibition of HIV-1 replication

Expression of outer capsid protein VP5 of grass carp reovirus in E.coli and analysis of its immunogenicity

HIV-1 Vif protein mediates the degradation of APOBEC3G in fission yeast when over-expressed using codon optimization

Genetic modification of baculovirus expression vectors

An improved strategy for efficient expression and purification of soluble HIV-1 tat protein in E.coli

Similar Documents

Characterization of synonymous codon usage bias in the pseudorabies virus US1 gene

Analysis of synonymous codon usage bias in 09H1N1

A new strategy for full-length Ebola virus glycoprotein expression in E.coli

Prokaryotic expression and purification of HIV-1 Vif and hAPOBEC3G, preparation of polyclonal antibodies

Synonymous codon usage bias and overexpression of a synthetic gene encoding Interferon α2b in yeast

Efficient gene transfer mediated by HIV-1-based defective lentivector and inhibition of HIV-1 replication

Expression of outer capsid protein VP5 of grass carp reovirus in E.coli and analysis of its immunogenicity

HIV-1 Vif protein mediates the degradation of APOBEC3G in fission yeast when over-expressed using codon optimization

Genetic modification of baculovirus expression vectors

An improved strategy for efficient expression and purification of soluble HIV-1 tat protein in E.coli