Studies on expression, purification, crystal growth and optimization of putative transcription factor LytR from Streptococcus pneumoniae / 生物医学工程学杂志
Journal of Biomedical Engineering
; (6): 812-821, 2013.
Article
in Zh
| WPRIM
| ID: wpr-352160
Responsible library:
WPRO
ABSTRACT
The aim of the present study was to obtain the crystal of transcription factor LytR of streptococcus pneumoniae for X-ray crystal structure and function analysis. The LytR gene of D39 strains from Streptococcus pneumoniae (S. pn) was cloned into the prokaryotic expression vector pET32a(+), then overexpression was obtained in the E. coli BL21 (DE3) through transformation of the recombinant plasmid that had been verified by colony PCR and sequencing. Soluble fusion protein with His-tag highly expressed by the induction of 0.5 mmol/L IPTG and was purified by a three step procedure, the purity of the purified LytR recombinant protein was over 90%. Preliminary screening of crystallization conditions was performed using the hanging-drop vapour-diffusing method with Hampton Crystal screen and PEG screen kits. The protein crystals X-ray diffraction data were collected from a single crystal and more stick crystals whose X-ray diffraction reached 4.0 A were obtained. These works laid the foundation for further research on the 3D structure of putative transcription factor LytR and its biological aspects.
Full text:
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Index:
WPRIM
Main subject:
Streptococcus pneumoniae
/
Bacterial Proteins
/
Transcription Factors
/
Recombinant Proteins
/
Escherichia coli
/
Genetics
/
Metabolism
Language:
Zh
Journal:
Journal of Biomedical Engineering
Year:
2013
Type:
Article