Expression of human pituitary adenylate cyclase activating polypeptide (PACAP) cDNA in CHO cells and characterization of the products

FEBS Letters

Okazaki, Kimitake¹  Onda, Haruo¹  Kitada, Chieko¹  Kimura, Chiharu¹  Ohkubo, Shoichi¹  Kosaka, Takuo¹  Ogi, Kazuhiro¹  Fujino, Masahiko¹  Watanabe, Takuya¹ 

[1] Tsukuba Research Laboratories, Takeda Chemical Industries Ltd., 10 Wadai, Tsukuba, Ibaraki 300-42, Japan

关键词: Human PACAP;    PRP;    Gene expression system (CHO-K1);    PC12h;    Neurite extension;    cAMP accumulation;    PACAP;    pituitary adenylate cyclase activating polypeptide;    PRP;    PACAP-related peptide;    ir-PACAP;    immunoreactive PACAP;    FBS;    fetal bovine serum;    HRP;    horseradish peroxidase;    EIA;    enzyme immunoassay;    FITC;    fluorescein isothiocyanate;    PBS;    phosphate-buffered saline;    HBSS;    Hanks' balanced salt solution;    TFA;    trifluoroacetic acid;    RP-HPLC;    reverse-phase high performance liquid chromatography;   

DOI  :  10.1016/0014-5793(92)80020-H

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

cDNA encoding human PACAP precursor was expressed in non-neuroendocrine Chinese hamster ovary cells, CHO-K1. The cells were transfected with expression vector (pTS705) containing the human PACAP cDNA by electroporation. A cell line which produced more than 80 ng/ml of immunoreactive PACAP (ir-PACAP) into the conditioned medium was established. RP-HPLC analysis of culture medium of this established cell line exhibited the presence of two types of PACAP, i.e. PACAP38 and PACAP27. At the same time, it was also revealed that immunoreactive PACAP-related peptide (ir-PRP) was secreted into the cultured medium. The ir-PACAPs were confirmed to have biological activities such as induction of cAMP and neurite outgrowth in rat pheochromocytoma PC12h cells.

【 授权许可】

Unknown