沙鼠白细胞介素12的分子克隆及其作为生物活性单链蛋白的表达

Molecular cloning of gerbil interleukin 12 and its expression as a bioactive single-chain protein.

作者信息

Gaucher D, Chadee K

机构信息

Institute of Parasitology, McGill University, Macdonald Campus, 21,111 Lakeshore Road, Ste. Anne de Bellevue, Quebec, Canada H9X 3V9.

出版信息

Cytokine. 2001 May 7;14(3):177-83. doi: 10.1006/cyto.2001.0866.

DOI:10.1006/cyto.2001.0866

PMID:11396996

Abstract

Complementary DNAs coding for gerbil interleukin 12 (IL-12) p40/p35 subunits were cloned by a combination of cross species reverse transcriptase-polymerase chain reaction (RT-PCR) and 3' rapid amplification of cDNA ends (RACE) techniques. IL-12 p40/p35 had 79% nucleotide identity and 81% amino acid homology to mouse IL-12 p40/p35. The p40/p35 subunits were expressed as a single polypeptide separated by a short hinge sequence that allowed for proper folding and assembly. COS-7 cells transfected with DNA encoding the single-chain gerbil IL-12 (pSCjIL12) secreted high levels of the protein which stimulated proliferation of ConA-activated gerbil spleen lymphoblasts in a dose-dependent manner.

摘要

通过跨物种逆转录聚合酶链反应（RT-PCR）和3' cDNA末端快速扩增（RACE）技术相结合，克隆了编码沙鼠白细胞介素12（IL-12）p40/p35亚基的互补DNA。IL-12 p40/p35与小鼠IL-12 p40/p35具有79%的核苷酸同一性和81%的氨基酸同源性。p40/p35亚基作为一个由短铰链序列分隔的单一多肽表达，该序列允许正确折叠和组装。用编码单链沙鼠IL-12（pSCjIL12）的DNA转染的COS-7细胞分泌高水平的蛋白质，该蛋白质以剂量依赖性方式刺激ConA激活的沙鼠脾脏淋巴母细胞增殖。