[Expression and purification of Rv3369 of Mycobacterium tuberculosis].
To obtain purified recombinant Rv3369 protein by means of expressing the Rv3369 protein of Mycobacterium tuberculosis in E. coli. The gene coding Rv3369 protein was amplified by polymerase chain reaction (PCR), then was inserted into an expression vector pET28a to get recombinant plasmid. The recombinant plasmid was transformed into E. coli BL21(DE3) and induced by IPTG. The expressed product was indentified by SDS-PAGE and purified by Ni- NTA His. Bind Resin. The sequence of Rv3369 in recombinant plasmid was the same with GenBank's report. The molecular mass of the product is 19.5kDa, which accounts for about 20% in the thalli proteins, and its purity is more than 90% analyzed by SDS-PAGE and laser scanning. The yield of recombinant protein is 1.56mg from 100mL of culture. Compared with other methods, purity of the recombinant protein is higher through affinity chromatography.
Lv
Chong
C
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China. lvchong0426@sohu.com
Jiang
Xin
X
Gu
Xiao-Ling
XL
Wang
Hong-Hai
HH
D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics*; D013058:Mass Spectrometry; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification
D001426:Bacterial Proteins; C117940:CFP-10 protein, Mycobacterium tuberculosis; D011994:Recombinant Proteins