人全长TALL-1在大肠杆菌中的表达

目的在克隆人TALL-1(TNF- and apoptosis ligand-related leukocyte expressed ligand 1)全长 cDNA的基础上,用大肠杆菌进行表达并鉴定其生物学活性.方法采用RT-PCR技术,从HL-60细胞中扩增编码人TALL-1的cDNA,克隆于高效原核表达载体pQE-80L中,以IPTG诱导表达,Ni-NTA层析纯化后,进行生物学活性检测.结果 RT-PCR扩增得到了858 bp的DNA片段,测序结果与GenBank中报道的编码TALL-1的cDNA序列一致,并在大肠杆菌中获得了高效表达,活性检测发现它能抑制U937细胞的生长.结论成功克隆了人TALL-1全长cDNA,并获得了高效表达及纯化,纯化产物具有生物学活性,这为进一步的功能研究及临床应用奠定了基础.

High expression of human TALL-1 in Escherichia coli

Objective To clone the cDNA encoding human TNF and apoptosis ligand related leukocyte expressed ligand 1 (TALL 1), to express it in Escherichia coli ( E. coli ), and to identify its biological activity. Methods The total RNA was extracted from HL 60 cells, and the cDNA encoding human TALL 1 amino acids was amplified by RT PCR. After sequence, the cDNA was ligated into the prokaryotic expression vector pQE 80L and induced by IPTG to express TALL 1. The protein was purified with Ni NTA chromatography, and its immunological activity was detected. Results The cDNA of 858 bp amplified by RT PCR was consistent with the sequence encoding human TALL 1 amino acids reported in GenBank. The TALL 1 was highly expressed. The protein could strongly suppress the growth of U937 cells. Conclusion The cDNA encoding human TALL 1 has been successfully cloned, and the relevant protein is highly expressed and purified. The purified product has biological activity. These results may pave the way for further studies.