Fusion protein of the hyaluronan binding domain from human TSG-6 with luciferase for assay of hyaluronan.
The gene expression plasmid, pET-Lmluc, for the fusion protein of the hyaluronan binding domain from human TSG-6 [product of tumor necrosis factor (TNF)-stimulated gene-6] and luciferase from Renilla reniformis was constructed. The fused gene was expressed in Escherichia coli and the resulted insoluble Lm-luc fusion protein was purified and refolded to recover both the hyaluronan binding capability and the luciferase activity. Hyaluronan as low as 1 ng ml(-1) was detected by using the indirect enzymatic immunological assay with the refolded Lm-luc fusion protein.
Chang
Te-Sheng
TS
Department of Chemical Engineering, National Tsing Hua University, Hsinchu 30043, Taiwan.
Wan
Hisao-Ming
HM
Chen
Chien-Chiang
CC
Giridhar
R
R
Wu
Wen-Teng
WT
D001665:Binding Sites; D015374:Biosensing Techniques / Q000379:methods*; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006820:Hyaluronic Acid / Q000032:analysis*; D007124:Immunoenzyme Techniques / Q000379:methods*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012680:Sensitivity and Specificity
D015815:Cell Adhesion Molecules; D011993:Recombinant Fusion Proteins; C072273:TNFAIP6 protein, human; D006820:Hyaluronic Acid; D008156:Luciferases