This study is conducted to clone the cDNA encoding human TNF-related apoptosis-inducing ligand (hTRAIL) extracellular region (amino acids 41-281, hTRAIL41-281) and to express it in E.coli. The hTRAIL41-281 cDNA is amp...This study is conducted to clone the cDNA encoding human TNF-related apoptosis-inducing ligand (hTRAIL) extracellular region (amino acids 41-281, hTRAIL41-281) and to express it in E.coli. The hTRAIL41-281 cDNA is amplified by reverse transcription (RT) PCR from total RNA derived from human acute promyelocytic leukemia cell line HL-60. After sequenced, the cDNA is cloned into the vector pQE-80L and transformed into E.coli DH5 to express the recombinant hTRAIL41-281 (rhTRAIL41-281) induced by IPTG. The recombinant protein is analyzed by SDS-PAGE. The cloned cDNA is consistent with the cDNA sequence encoding hTRAIL41-281 reported in GenBankTM. After inducing, the hTRAIL41-281 protein is expressed, and the mass of the recombinant protein is about 30 % of total bacteria protein, which demonstrates that the cDNA encoding hTRAIL41-281 is successfully cloned and expressed in E.coli.展开更多
基金Funded by the Scientific Research Foundation of the Third Military Medical University.
文摘This study is conducted to clone the cDNA encoding human TNF-related apoptosis-inducing ligand (hTRAIL) extracellular region (amino acids 41-281, hTRAIL41-281) and to express it in E.coli. The hTRAIL41-281 cDNA is amplified by reverse transcription (RT) PCR from total RNA derived from human acute promyelocytic leukemia cell line HL-60. After sequenced, the cDNA is cloned into the vector pQE-80L and transformed into E.coli DH5 to express the recombinant hTRAIL41-281 (rhTRAIL41-281) induced by IPTG. The recombinant protein is analyzed by SDS-PAGE. The cloned cDNA is consistent with the cDNA sequence encoding hTRAIL41-281 reported in GenBankTM. After inducing, the hTRAIL41-281 protein is expressed, and the mass of the recombinant protein is about 30 % of total bacteria protein, which demonstrates that the cDNA encoding hTRAIL41-281 is successfully cloned and expressed in E.coli.